{ "Contributors": "HoC", "Source": "HoC", "URL": "https://github.com/sb895/Hallmarks-of-Cancer", "Categories": [ "Text Classification" ], "Definition": [ "Given the biomedical publication abstracts, you are required to classify the hallmarks of cancer. Each article may have one or more of the ten hallmarks. The ten hallmarks are as follows: 1. Sustaining proliferative signaling, 2. Evading growth suppressors, 3. Resisting cell death, 4. Enabling replicative immortality, 5. Inducing angiogenesis, 6. Activating invasion and metastasis, 7. Genomic instability and mutation, 8. Tumor promoting inflammation, 9. Cellular energetics, 10. Avoiding immune destruction." ], "Reasoning": [], "Input_language": [ "English" ], "Output_language": [ "English" ], "Instruction_language": [ "English" ], "Domains": [ "Public Health", "Heathcare" ], "Positive Examples": [], "Negative Examples": [], "Instances": [ { "input": "MicroRNAs ( miRNAs ) are involved in cancer development and progression , acting as tumor suppressors or oncogenes .\n\nIn this study , miRNA profiling was performed on 10 paired bladder cancer ( BC ) tissues using 20 GeneChipTM miRNA Array , and 10 differentially expressed miRNAs were identified in BC and adjacent noncancerous tissues of any disease stage/grade .\n\nAfter validated on expanded cohort of 67 paired BC tissues and 10 human BC cell lines by qRT-PCR , it was found that miR-100 was down-regulated most significantly in cancer tissues .\n\nEctopic restoration of miR-100 expression in BC cells suppressed cell proliferation and motility , induced cell-cycle arrest in vitro , and inhibited tumorigenesis in vivo both in subcutaneous and intravesical passage .\n\nBioinformatic analysis showed that mTOR gene was a direct target of miR-100. siRNA-mediated mTOR knockdown phenocopied the effect of miR-100 in BC cell lines .\n\nIn addition , the cancerous metastatic nude mouse model established on the basis of primary BC cell lines suggested that miR-100/mTOR regulated cell motility and was associated with tumor metastasis .\n\nBoth mTOR and p70S6K ( downstream messenger ) presented higher expression levels in distant metastatic foci such as in liver and kidney metastases than in primary tumor .\n\nTaken together , miR-100 may act as a tumor suppressor in BC , and reintroduction of this mature miRNA into tumor tissue may prove to be a therapeutic strategy by reducing the expression of target genes .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Activating invasion and metastasis" }, { "input": "Diffuse intrinsic pontine glioma ( DIPG ) is a fatal pediatric disease .\n\nThus far no therapeutic agent has proven beneficial in the treatment of this malignancy .\n\nHence , conventional DNA-damaging radiotherapy ( RT ) remains the standard treatment , providing transient neurological improvement without improving probability of overall survival .\n\nDuring RT , WEE1 kinase controls the G2 cell cycle checkpoint allowing for repair of irradiation ( IR)-induced DNA damage .\n\nHere we show that WEE1 kinase is one of the highest overexpressed kinases in primary DIPG tissues as compared to matching non-neoplastic brain tissues .\n\nInhibition of WEE1 by MK-1775 treatment of DIPG cells inhibited the IR-induced WEE1-mediated phosphorylation of CDC2 , resulting in reduced G2/M arrest and decreased cell viability .\n\nFinally , we demonstrate that MK-1775 enhances the radiation response of E98-Fluc-mCherry DIPG mouse xenografts .\n\nAltogether , these results show that inhibition of WEE1 kinase in conjunction with RT holds potential as a therapeutic approach for the treatment of DIPG .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "OBJECTIVES : Cigarette smoking is a major risk factor for pancreatic cancer ( PaCa ) .\n\nHowever , the mechanisms of smoking-induced PaCa remain unknown .\n\nHere we investigated the effect of smoking compounds on cell death pathways in pancreatic ductal cells , precursors of PaCa .\n\nMETHODS : Human pancreatic ductal cells ( HPDE6-c7 ) were cultured with cigarette smoke extract ( CSE ) or smoking compound 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone ( NNK ) .\n\nApoptosis and autophagy were assessed by DNA fragmentation and immunofluorescence , respectively .\n\nRESULTS : Exposure to CSE or NNK decreased DNA fragmentation and up-regulated BclXL .\n\nAkt kinase was activated by smoking compounds through reactive oxygen species-dependent mechanism .\n\nSpecifically , Akt activation was prevented by inhibition of nicotinamide adenine dinucleotide oxidase .\n\nMolecular or pharmacologic inhibitions of Akt prevented the antiapoptotic effect of smoking compounds .\n\nSmoking compounds stimulated rapid ( 1 hour ) and transient activation of 5'-adenosine monophosphate-activated protein kinase and formation of autophagic vacuoles , indicating stimulation of autophagy .\n\nRepeated exposure to CSE/NNK ( 48 hours or longer ) abolished the early activation of autophagic markers .\n\nInhibition of Akt prevented the antiautophagic effect of long exposure to smoking compounds , indicating that smoking-induced late activation of Akt prevents autophagy .\n\nCONCLUSIONS : Long exposure of pancreatic ductal cells to smoking compounds inhibited apoptosis and autophagy .\n\nThe results revealed a central role for Akt kinase in mediating key procarcinogenic effects of smoking compounds .", "output": "Tumor promoting inflammation, Resisting cell death" }, { "input": "Most hepatocellular carcinomas ( HCC ) develop in the context of severe liver fibrosis and cirrhosis caused by chronic liver inflammation , which also results in accumulation of reactive oxygen species ( ROS ) .\n\nIn this study , we examined whether the stress-activated protein kinase p38\u03b1 ( Mapk14 ) controls ROS metabolism and development of fibrosis and cancer in mice given thioacetamide to induce chronic liver injury .\n\nLiver-specific p38\u03b1 ablation was found to enhance ROS accumulation , which appears to be exerted through the reduced expression of antioxidant protein HSP25 ( Hspb1 ) , a mouse homolog of HSP27 .\n\nIts reexpression in p38\u03b1-deficient liver prevents ROS accumulation and thioacetamide-induced fibrosis. p38\u03b1 deficiency increased expression of SOX2 , a marker for cancer stem cells and the liver oncoproteins c-Jun ( Jun ) and Gankyrin ( Psmd10 ) and led to enhanced thioacetamide-induced hepatocarcinogenesis .\n\nThe upregulation of SOX2 and c-Jun was prevented by administration of the antioxidant butylated hydroxyanisole .\n\nIntriguingly , the risk of human HCC recurrence is positively correlated with ROS accumulation in liver .\n\nThus , p38\u03b1 and its target HSP25/HSP27 appear to play a conserved and critical hepatoprotective function by curtailing ROS accumulation in liver parenchymal cells engaged in oxidative metabolism of exogenous chemicals .\n\nAugmented oxidative stress of liver parenchymal cells may explain the close relationship between liver fibrosis and hepatocarcinogenesis .", "output": "Tumor promoting inflammation" }, { "input": "The role of inflammatory cytokine interleukin-20 ( IL-20 ) has not yet been studied in cancer biology .\n\nHere , we demonstrated up-regulation of both IL-20 and IL-20R1 in muscle invasive bladder cancer ( MIBC ) patients .\n\nThe expressions of IL-20 and IL-20R1 were observed in bladder cancer 5637 and T-24 cells .\n\nWe found that IL-20 significantly increased the expression of matrix metalloproteinase ( MMP)-9 via binding activity of NF-\u03baB and AP-1 in bladder cancer cells and stimulated the activation of ERK1/2 , JNK , p38MAPK , and Jak-Stat signaling .\n\nAmong the pathways examined , only ERK1/2 inhibitor U0126 significantly inhibited IL-20-induced migration and invasion .\n\nMoreover , siRNA knockdown of IL-20R1 suppressed migration , invasion , ERK1/2 activation , and NF-\u03baB-mediated MMP-9 expression induced by IL-20 .\n\nUnexpectedly , cell cycle inhibitor p21WAF1 was induced by IL-20 treatment without altering cell cycle progression .\n\nBlockade of p21WAF1 function by siRNA reversed migration , invasion , activation of ERK signaling , MMP-9 expression , and activation of NF-\u03baB in IL-20-treated cells .\n\nIn addition , IL-20 induced the activation of IKK , the degradation and phosphorylation of I\u03baBa , and NF-\u03baB p65 nuclear translocation , which was regulated by ERK1/2 .\n\nIL-20 stimulated the recruitment of p65 to the MMP-9 promoter region .\n\nFinally , the IL-20-induced migration and invasion of cells was confirmed by IL-20 gene transfection and by addition of anti-IL-20 antibody .\n\nThis is the first report that p21WAF1 is involved in ERK1/2-mediated MMP-9 expression via increased binding activity of NF-\u03baB , which resulted in the induction of migration in IL-20/IL-20R1 dyad-induced bladder cancer cells .\n\nThese unexpected results might provide a critical new target for the treatment of bladder cancer .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Activating invasion and metastasis" }, { "input": "Each year , more than 700,000 people undergo cancer surgery in the United States .\n\nHowever , more than 40% of those patients develop recurrences and have a poor outcome .\n\nTraditionally , the medical community has assumed that recurrent tumors arise from selected tumor clones that are refractory to therapy .\n\nHowever , we found that tumor cells have few phenotypical differences after surgery .\n\nThus , we propose an alternative explanation for the resistance of recurrent tumors .\n\nSurgery promotes inhibitory factors that allow lingering immunosuppressive cells to repopulate small pockets of residual disease quickly .\n\nRecurrent tumors and draining lymph nodes are infiltrated with M2 ( CD11b(+)F4/80(hi)CD206(hi) and CD11b(+)F4/80(hi)CD124(hi) ) macrophages and CD4(+)Foxp3(+) regulatory T cells .\n\nThis complex network of immunosuppression in the surrounding tumor microenvironment explains the resistance of tumor recurrences to conventional cancer vaccines despite small tumor size , an intact antitumor immune response , and unaltered cancer cells .\n\nTherapeutic strategies coupling antitumor agents with inhibition of immunosuppressive cells potentially could impact the outcomes of more than 250,000 people each year .", "output": "Avoiding immune destruction" }, { "input": "Chromosomal DNA must be in single-strand form for important transactions such as replication , transcription , and recombination to occur .\n\nThe single-strand DNA ( ssDNA ) is more prone to damage than double-strand DNA ( dsDNA ) , due to greater exposure of chemically reactive moieties in the nitrogenous bases .\n\nThus , there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA .\n\nTo assess the potential hazard posed by such agents , we devised an ssDNA-specific mutagenesis reporter system in budding yeast .\n\nThe reporter strains bear the cdc13-1 temperature-sensitive mutation , such that shifting to 37\u00b0C results in telomere uncapping and ensuing 5 ' to 3 ' enzymatic resection .\n\nThis exposes the reporter region , containing three closely-spaced reporter genes , as a long 3 ' ssDNA overhang .\n\nWe validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase , APOBEC3G .\n\nAPOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand , resulting in frequent , simultaneous inactivation of two reporter genes .\n\nWe then examined the mutagenicity of sulfites , a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake .\n\nSulfites , at a concentration similar to that found in some foods , induced a high density of mutations , almost always as substitutions at cytosines in the ssDNA overhang strand , resulting in simultaneous inactivation of at least two reporter genes .\n\nFurthermore , sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase .\n\nThis intermediate was bypassed by error-prone translesion DNA synthesis , frequently involving Pol \\u03b6 , during repair synthesis .\n\nOur results suggest that sulfite-induced lesions in DNA can be particularly deleterious , since cells might not possess the means to repair or bypass such lesions accurately .", "output": "Genomic instability and mutation" }, { "input": "Hypoxia is known to play critical roles in cell survival , angiogenesis , tumor invasion , and metastasis .\n\nHypoxia mediated over-expression of hypoxia-inducible factor ( HIF ) has been shown to be associated with therapeutic resistance , and contributes to poor prognosis of cancer patients .\n\nEmerging evidence suggest that hypoxia and HIF pathways contributes to the acquisition of epithelial-to-mesenchymal transition ( EMT ) , maintenance of cancer stem cell ( CSC ) functions , and also maintains the vicious cycle of inflammation-all which lead to therapeutic resistance .\n\nHowever , the precise molecular mechanism(s) by which hypoxia/HIF drives these events are not fully understood .\n\nHere , we show , for the first time , that hypoxia leads to increased expression of VEGF , IL-6 , and CSC signature genes Nanog , Oct4 and EZH2 consistent with increased cell migration/invasion and angiogenesis , and the formation of pancreatospheres , concomitant with increased expression of miR-21 and miR-210 in human pancreatic cancer ( PC ) cells .\n\nThe treatment of PC cells with CDF , a novel synthetic compound inhibited the production of VEGF and IL-6 , and down-regulated the expression of Nanog , Oct4 , EZH2 mRNAs , as well as miR-21 and miR-210 under hypoxia .\n\nCDF also led to decreased cell migration/invasion , angiogenesis , and formation of pancreatospheres under hypoxia .\n\nMoreover , CDF decreased gene expression of miR-21 , miR-210 , IL-6 , HIF-1\u03b1 , VEGF , and CSC signatures in vivo in a mouse orthotopic model of human PC .\n\nCollectively , these results suggest that the anti-tumor activity of CDF is in part mediated through deregulation of tumor hypoxic pathways , and thus CDF could become a novel , and effective anti-tumor agent for PC therapy .", "output": "Inducing angiogenesis, Activating invasion and metastasis" }, { "input": "The ATM/ATR DNA damage checkpoint functions in the maintenance of genetic stability and some missense variants of the ATM gene have been shown to confer a moderate increased risk of prostate cancer .\n\nHowever , whether inactivation of this checkpoint contributes directly to prostate specific cancer predisposition is still unknown .\n\nHere , we show that exposure of non-malignant prostate epithelial cells ( HPr-1AR ) to androgen led to activation of the ATM/ATR DNA damage response and induction of cellular senescence .\n\nNotably , knockdown of the ATM gene expression in HPr-1AR cells can promote androgen-induced TMPRSS2 : ERG rearrangement , a prostate-specific chromosome translocation frequently found in prostate cancer cells .\n\nIntriguingly , unlike the non-malignant prostate epithelial cells , the ATM/ATR DNA damage checkpoint appears to be defective in prostate cancer cells , since androgen treatment only induced a partial activation of the DNA damage response .\n\nThis mechanism appears to preserve androgen induced autophosphorylation of ATM and phosphorylation of H2AX , lesion processing and repair pathway yet restrain ATM/CHK1/CHK2 and p53 signaling pathway .\n\nOur findings demonstrate that ATM/ATR inactivation is a crucial step in promoting androgen-induced genomic instability and prostate carcinogenesis .", "output": "Genomic instability and mutation, Enabling replicative immortality" }, { "input": "Human CD93 , an epidermal growth factor ( EGF)-like domain containing transmembrane protein , is predominantly expressed in the vascular endothelium .\n\nStudies have shown that AA4 , the homolog of CD93 in mice , may mediate cell migration and angiogenesis in endothelial cells .\n\nSoluble CD93 has been detected in the plasma of healthy individuals .\n\nHowever , the role of soluble CD93 in the endothelium remains unclear .\n\nRecombinant soluble CD93 proteins with EGF-like domains ( rCD93D123 , with domains 1 , 2 , and 3 ; and rCD93D23 , with domains 2 and 3 ) were generated to determine their functions in angiogenesis .\n\nWe found that rCD93D23 was more potent than rCD93D123 in stimulating the proliferation and migration of human umbilical vein endothelial cells ( HUVECs ) .\n\nProduction of matrix-metalloproteinase 2 increased after the HUVECs were treated with rCD93D23 .\n\nFurther , in a tube formation assay , rCD93D23 induced cell differentiation of HUVECs through phosphoinositide 3-kinase/Akt/endothelial nitric oxide synthase and extracellular signal-regulated kinases-1/2 signaling .\n\nMoreover , rCD93D23 promoted blood vessel formation in a Matrigel-plug assay and an oxygen-induced retinopathy model in vivo .\n\nOur findings suggest that the soluble EGF-like domain containing CD93 protein is a novel angiogenic factor acting on the endothelium .", "output": "Inducing angiogenesis" }, { "input": "Prostate cancer is the most frequent and second most lethal cancer in men in the United States .\n\nInnate immunity and inflammation may increase the risk of prostate cancer .\n\nTo determine the role of innate immunity and inflammation in advanced prostate cancer , we investigated the association of 320 single nucleotide polymorphisms , located in 46 genes involved in this pathway , with disease risk using 494 cases with advanced disease and 536 controls from Cleveland , Ohio .\n\nTaken together , the whole pathway was associated with advanced prostate cancer risk ( P\u200a=\u200a0.02 ) .\n\nTwo sub-pathways ( intracellular antiviral molecules and extracellular pattern recognition ) and four genes in these sub-pathways ( TLR1 , TLR6 , OAS1 , and OAS2 ) were nominally associated with advanced prostate cancer risk and harbor several SNPs nominally associated with advanced prostate cancer risk .\n\nOur results suggest that the innate immunity and inflammation pathway may play a modest role in the etiology of advanced prostate cancer through multiple small effects .", "output": "Tumor promoting inflammation" }, { "input": "Ulcerative colitis ( UC ) is a major form of chronic inflammation that can frequently progress to colon cancer .\n\nSeveral studies have demonstrated massive infiltration of neutrophils and macrophages into the lamina propria and submucosa in the progression of UC-associated colon carcinogenesis .\n\nMacrophages contribute to the development of colitis-associated colon cancer ( CAC ) .\n\nHowever , the role of neutrophils is not well understood .\n\nTo better understand the involvement of tumor-associated neutrophils ( TANs ) in the regulation of CAC , we used a mouse CAC model produced by administering azoxymethane ( AOM ) , followed by repeated dextran sulfate sodium ( DSS ) ingestion .\n\nThis causes severe colonic inflammation and subsequent development of multiple tumors in mice colon .\n\nWe observed that colorectal mucosal inflammation became increasingly severe with AOM and DSS treatment .\n\nMacrophages infiltrated the lamina propria and submucosa , together with a marked increase in neutrophil infiltration .\n\nThe chemokine CXCL2 increased in the lamina propria and submucosal regions of the colons of the treated mice , together with the infiltration of neutrophils expressing CXCR2 , a specific receptor for CXCL2 .\n\nThis process was followed by neoplastic transformation .\n\nAfter AOM and DSS treatment , the mice showed enhanced production of metalloproteinase ( MMP)-9 and neutrophil elastase ( NE ) , accompanied by excessive vessel generation and cell proliferation .\n\nMoreover , CXCL2 promoted neutrophil recruitment and induced neutrophils to express MMP-9 and NE in vitro .\n\nFurthermore , administration of neutrophil-neutralizing antibodies after the last DSS cycle markedly reduced the number and size of tumors and decreased the expression of CXCR2 , CXCL2 , MMP-9 , and NE .\n\nThese observations indicate a crucial role for TANs in the initiation and progression of CAC and suggest that the CXCL2-CXCR2 axis might be useful in reducing the risk of UC-associated colon cancer .", "output": "Inducing angiogenesis, Tumor promoting inflammation" }, { "input": "Cellular senescence is considered as a tumor suppressive mechanism .\n\nRecent evidence indicates however that senescent cells secrete various growth factors and cytokines , some of which may paradoxically promote cancer progression .\n\nThis phenomenon termed senescence-associated secretory phenotype ( SASP ) must be inhibited in order for anti-proliferative agents to be effective .\n\nThe present study was designed to determine whether the \u03b2-catenin destruction complex ( BCDC ) , known to integrate the action of various growth factors and cytokines , would represent a suitable target to inhibit the activity of SASP components .\n\nFor this , we carried out experiments to determine the effect of drug-induced senescence on secretion of SASP , \u03b2-catenin transactivation , and the relationship between these processes .\n\nMoreover , genetic and pharmacological approaches were used to define the implication of BCDC in mediating the effects of SASP components on cell migration and resistance to drugs .\n\nThe findings indicate that drug-induced senescence was associated with expression of various Wnt ligands in addition to previously known SASP components .\n\nBeta catenin transactivation and expression of genes implicated in epithelial-mesenchymal transition ( EMT ) also increased in response to drug-induced SASP .\n\nThese effects were prevented by Pyrvinium , a recently described activator of BCDC .\n\nPyrvinium also suppressed the effects of SASP on cell migration and resistance to doxorubicin .\n\nTogether , these findings provide insights on the potential role of BCDC in mediating the effects of drug-induced SASP on cancer cell invasion and resistance to therapy , and suggest that targeting this pathway may represent an effective approach to enhance the activity of current and prospective anti-cancer therapeutics .", "output": "Enabling replicative immortality, Activating invasion and metastasis" }, { "input": "Loss of p53 is considered to allow progression of colorectal tumors from the adenoma to the carcinoma stage .\n\nUsing mice with an intestinal epithelial cell ( IEC)-specific p53 deletion , we demonstrate that loss ofp53 alone is insufficient to initiate intestinal tumorigenesis but markedly enhances carcinogen-induced tumor incidence and leads to invasive cancer and lymph node metastasis .\n\nWhereas p53 controls DNA damage and IEC survival during the initiation stage , loss of p53 during tumor progression is associated with increased intestinal permeability , causing formation of an NF-\u03baB-dependent inflammatory microenvironment and the induction of epithelial-mesenchymal transition .\n\nThus , we propose a p53-controlled tumor-suppressive function that is independent of its well-established role in cell-cycle regulation , apoptosis , and senescence .", "output": "Enabling replicative immortality, Activating invasion and metastasis, Evading growth suppressors, Tumor promoting inflammation, Resisting cell death" }, { "input": "Binding within or nearby target genes involved in cell proliferation and survival enables the p53 tumor suppressor gene to regulate their transcription and cell-cycle progression .\n\nUsing genome-wide chromatin-binding profiles , we describe binding of p53 also to regions located distantly from any known p53 target gene .\n\nInterestingly , many of these regions possess conserved p53-binding sites and all known hallmarks of enhancer regions .\n\nWe demonstrate that these p53-bound enhancer regions ( p53BERs ) indeed contain enhancer activity and interact intrachromosomally with multiple neighboring genes to convey long-distance p53-dependent transcription regulation .\n\nFurthermore , p53BERs produce , ina p53-dependent manner , enhancer RNAs ( eRNAs ) that are required for efficient transcriptional enhancement of interacting target genes and induction of a p53-dependent cell-cycle arrest .\n\nThus , our results ascribe transcription enhancement activity to p53 with the capacity to regulate multiple genes from a single genomic binding site .\n\nMoreover , eRNA production from p53BERs is required for efficient p53 transcription enhancement .", "output": "Evading growth suppressors" }, { "input": "Jaceosidin , a flavonoid derived from Artemisia princeps ( Japanese mugwort ) , has been shown to inhibit the growth of several human cancer cells , However , the exact mechanism for the cytotoxic effect of jaceosidin is not completely understood .\n\nIn this study , we investigated the molecular mechanism involved in the antiproliferative effect of jaceosidin in human endometrial cancer cells .\n\nWe demonstrated that jaceosidin is a more potent inhibitor of cell growth than cisplatin in human endometrial cancer cells .\n\nIn contrast , jaceosidin-induced cytotoxicity in normal endometrial cells was lower than that observed for cisplatin .\n\nJaceosidin induced G2/M phase cell cycle arrest and modulated the levels of cyclin B and p-Cdc2 in Hec1A cells .\n\nKnockdown of p21 using specific siRNAs partially abrogated jaceosidin-induced cell growth inhibition .\n\nAdditional mechanistic studies revealed that jaceosidin treatment resulted in an increase in phosphorylation of Cdc25C and ATM-Chk1/2 .\n\nKu55933 , an ATM inhibitor , reversed jaceosidin-induced cell growth inhibition , in part .\n\nMoreover , jaceosidin treatment resulted in phosphorylation of ERK , and pretreatment with the ERK inhibitor , PD98059 , attenuated cell growth inhibition by jaceosidin .\n\nThese data suggest that jaceosidin , isolated from Japanese mugwort , modulates the ERK/ATM/Chk1/2 pathway , leading to inactivation of the Cdc2-cyclin B1 complex , followed by G2/M cell cycle arrest in endometrial cancer cells .", "output": "Sustaining proliferative signaling" }, { "input": "AMPK is a metabolic sensor that helps maintain cellular energy homeostasis .\n\nDespite evidence linking AMPK with tumor suppressor functions , the role of AMPK in tumorigenesis and tumor metabolism isunknown .\n\nHere we show that AMPK negatively regulates aerobic glycolysis ( the Warburg effect ) in cancer cells and suppresses tumor growth invivo .\n\nGenetic ablation of the \\u03b11 catalytic subunit of AMPK accelerates Myc-induced lymphomagenesis .\n\nInactivation of AMPK\\u03b1 in both transformed and nontransformed cells promotes a metabolic shift to aerobic glycolysis , increased allocation of glucose carbon into lipids , and biomass accumulation .\n\nThese metabolic effects require normoxic stabilization of the hypoxia-inducible factor-1\\u03b1 ( HIF-1\\u03b1 ) , as silencing HIF-1\\u03b1 reverses the shift to aerobic glycolysis and the biosynthetic and proliferative advantages conferred by reduced AMPK\\u03b1 signaling .\n\nTogether our findings suggest that AMPK activity opposes tumor development and that its loss fosters tumor progression in part by regulating cellular metabolic pathways that support cell growth and proliferation .", "output": "Cellular energetics" }, { "input": "Indolylmaleimides display a broad spectrum of biological activity and offer great opportunity to influence several aspects of cell fate , as proliferation and differentiation .\n\nIn this study we describe the effect of PDA-66 , a newly synthesised indolylmaleimide , showing a strong dose dependent anti-proliferative effect on immortalised human progenitor and cancer cells .\n\nWe demonstrated a highly depolymerizing effect on in vitro tubulin assembly and conclude that PDA-66 acts as microtubule destabilising agent .\n\nIn addition we found that PDA-66 induces mitotic arrest of cells in the G(2)/M phase of the cell cycle .\n\nSubsequently cells undergo apoptosis , indicating the major mechanism of the anti-proliferative effect .\n\nTo prove a potential anti-cancer activity of PDA-66 we examined the effect of PDA-66 on human SH-SY5Y neuroblastoma and A-459 lung cancer cells , showing a significant reduction in cancer cell proliferation in a dose dependent manner .\n\nThus PDA-66 is a new anti-mitotic compound with an indole-core with the potential to be used for cancer therapy .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "In human epithelial cancers , the microRNA ( miRNA ) mir-30d is amplified with high frequency and serves as a critical oncomir by regulating metastasis , apoptosis , proliferation , and differentiation .\n\nAutophagy , a degradation pathway for long-lived protein and organelles , regulates the survival and death of many cell types .\n\nIncreasing evidence suggests that autophagy plays an important function in epithelial tumor initiation and progression .\n\nUsing a combined bioinformatics approach , gene set enrichment analysis and miRNA target prediction , we found that mir-30d might regulate multiple genes in the autophagy pathway including BECN1 , BNIP3L , ATG12 , ATG5 , ATG2 .\n\nOur further functional experiments demonstrated that the expression of these core proteins in the autophagy pathway was directly suppressed by mir-30d in cancer cells .\n\nFinally , we showed that mir-30d regulated the autophagy process by inhibiting autophagosome formation and LC3B-I conversion to LC3B-II .\n\nTaken together , our results provide evidence that the oncomir mir-30d impairs the autophagy process by targeting multiple genes in the autophagy pathway .\n\nThis result will contribute to understanding the molecular mechanism of mir-30d in tumorigenesis and developing novel cancer therapy strategy .", "output": "Resisting cell death" }, { "input": "Compound C , a well-known inhibitor of the intracellular energy sensor AMP-activated protein kinase ( AMPK ) , has been reported to cause apoptotic cell death in myeloma , breast cancer cells and glioma cells .\n\nIn this study , we have demonstrated that compound C not only induced autophagy in all tested skin cancer cell lines but also caused more apoptosis in p53 wildtype skin cancer cells than in p53-mutant skin cancer cells .\n\nCompound C can induce upregulation , phosphorylation and nuclear translocalization of the p53 protein and upregulate expression of p53 target genes in wildtype p53-expressing skin basal cell carcinoma ( BCC ) cells .\n\nThe changes of p53 status were dependent on DNA damage which was caused by compound C induced reactive oxygen species ( ROS ) generation and associated with activated ataxia-telangiectasia mutated ( ATM ) protein .\n\nUsing the wildtype p53-expressing BCC cells versus stable p53-knockdown BCC sublines , we present evidence that p53-knockdown cancer cells were much less sensitive to compound C treatment with significant G2/M cell cycle arrest and attenuated the compound C-induced apoptosis but not autophagy .\n\nThe compound C induced G2/M arrest in p53-knockdown BCC cells was associated with the sustained inactive Tyr15 phosphor-Cdc2 expression .\n\nOverall , our results established that compound C-induced apoptosis in skin cancer cells was dependent on the cell's p53 status .", "output": "Genomic instability and mutation, Sustaining proliferative signaling, Evading growth suppressors, Tumor promoting inflammation, Resisting cell death" }, { "input": "Bisphenol A ( BPA ) is one of the most prevalent chemicals in daily-use materials , therefore , human exposure to BPA is ubiquitous .\n\nWe found that low concentrations of BPA stimulate the spermatogonial GC-1 cells proliferation by G protein-coupled receptor 30 ( GPR30)-mediated epidermal growth factor receptor ( EGFR)-extracellular regulated kinase ( ERK)-c-Fos pathway .\n\nHowever , through the same pathway GPR30 expression has been shown to be induced by EGF , an EGFR ligand .\n\nThus , we want to know if low concentrations of BPA are able to induce the GPR30 expression and the possible mechanism(s) in GC-1 cells .\n\nBy transient transfection with expression plasmids , 10(-9)M BPA significantly transactivates the Gpr30-5'-flanking region through activating the GPR30 , cGMP-dependent protein kinase ( PKG ) , estrogen receptor-\u03b1 ( ER-\u03b1 ) , and EFGR-ERK pathways .\n\nFurthermore , an activator protein-1 ( AP-1 ) site located within this region is found to be responsible for the transactivation of BPA .\n\nExpectedly , through the same pathways , BPA significantly induces the gene and protein expression of GPR30. c-Fos is further observed to be strongly recruited to the AP-1 site in a chromatin immunoprecipitation assay and its dysfunction on the AP-1 site markedly suppresses the expression of GPR30 , p-ERK1/2 , p-Ser118-ER-\u03b1 and cell proliferation by BPA .\n\nOur results demonstrate that a low-concentration BPA induces GPR30 expression through the GPR30-EFGR-ERK-c-Fos , ER-\u03b1 , and PKG pathways , presumably boosting the cells proliferation via a regulatory loop .\n\nThe present study provides a novel insight into the potential role of GPR30 in the initiation and progression of male germ cell cancer induced by environmentally relevant BPA .", "output": "Sustaining proliferative signaling" }, { "input": "Nitric oxide ( NO)-releasing non-steroidal anti-inflammatory drugs ( NO-NSAIDs ) which have been synthesized to reduce gastro-intestinal and cardiovascular toxicities of NSAIDs , possess anti-proliferative , pro-apoptotic and anti-cancer activities .\n\nHere , we show that NO-sulindac inhibited UVB-induced skin tumorigenesis in SKH-1 hairless mice .\n\nTopical application of NO-sulindac reduced tumor incidence , number ( p<0.05 ) and volume ( p<0.005 ) as compared to UVB ( alone)-irradiated vehicle-treated mice .\n\nAn increase in TUNEL-positive cells in skin lesions was accompanied by the enhanced Bax:Bcl-2 ratio .\n\nThe expression of pro-apoptotic Bax was increased whereas anti-apoptotic Bcl-2 reduced .\n\nHowever , proliferation was identified as the major target of NO-sulindac in this study .\n\nA reduced expression of PCNA and cyclin D1 associated with the dampening of cell cycle progression was observed .\n\nThe mechanism of this inhibition was related to the reduction in UVB-induced Notch signaling pathway .\n\nUVB-induced inflammatory responses were diminished by NO-sulindac as observed by a remarkable reduction in the levels of phosphorylated MAP Kinases Erk1/2 , p38 and JNK1/2 .\n\nIn this regard , NO-sulindac also inhibited NF\u03baB by enhancing I\u03baB\u03b1 as evidenced by the reduced expression of iNOS and COX-2 , the direct NF\u03baB transcription target proteins .\n\nNO-sulindac significantly diminished the progression of benign lesions to invasive carcinomas by suppressing the tumor aggressiveness and retarding epithelial-mesenchymal transition .\n\nA marked decrease in the expression of mesenchymal markers such as Fibronectin , N-cadherin , SNAI , Slug and Twist and an increase in epithelial cell polarity marker E-cadherin were noted in NO-sulindac-treated tumors .\n\nOur data suggest that NO-sulindac is a potent inhibitor of UVB-induced skin carcinogenesis and acts by targeting proliferation-regulatory pathways .", "output": "Activating invasion and metastasis, Tumor promoting inflammation, Sustaining proliferative signaling, Resisting cell death" }, { "input": "Breast cancer is the second most common cancer with a high incidence rate worldwide .\n\nOne of the promising therapeutic approaches on breast cancer is to use the drugs that target the estrogen receptor ( ER ) .\n\nIn the present investigation , marmorin , a type I ribosome inactivating protein from the mushroom Hypsizigus marmoreus , inhibited the survival of breast cancer in vitro and in vivo .\n\nIt evinced more potent cytotoxicity toward estrogen receptor ( ER)-positive MCF7 breast cancer cells than ER-negative MDA-MB-231 cells .\n\nFurther study disclosed that marmorin undermined the expression level of estrogen receptor \u03b1 ( ER\u03b1 ) and significantly inhibited the proliferation of MCF7 cells induced by 17\u03b2-estradiol .\n\nKnockdown of ER\u03b1 in MCF7 cells significantly attenuated the inhibitory effect of marmorin on proliferation , suggesting that the ER\u03b1-mediated pathway was implicated in the suppressive action of marmorin on ER-positive breast cancer cells .\n\nMoreover , marmorin induced time-dependent apoptosis in both MCF7 and MDA-MB-231 cells .\n\nIt brought about G2/M-phase arrest , mitochondrial membrane potential depolarization and caspase-9 activation in MCF7 cells , and to a lesser extent in MDA-MB-231 cells .\n\nMarmorin triggered the death receptor apoptotic pathway ( e.g. caspase-8 activation ) and endoplasmic reticulum stress ( ERS , as evidenced by phosphorylation of PERK and IRE1\u03b1 , cleavage of caspase-12 , and up-regulation of CHOP expression ) in both MCF7 and MDA-MB-231 cells .\n\nIn summary , marmorin exhibited inhibitory effect on breast cancer partially via diminution of ER\u03b1 and apoptotic pathways mediated by mitochondrial , death receptor and ERS .\n\nThe results advocate that marmorin is a potential candidate for breast cancer therapy .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "A new line of human ovarian serous adenocarcinoma cells , TU-OS-4 , was established and characterized .\n\nThe cells showed a short , spindle-shaped morphology and grew in monolayers without contact inhibition while forming an arrangement resembling a jigsaw puzzle .\n\nChromosome numbers ranged from 55 to 73 .\n\nThe proliferation rate was lower than other serous adenocarcinoma cell lines tested ( KF , SHIN-3 , and SK-OV-3 ) , and the doubling time was 53.3 h .\n\nWestern blot analysis showed that TU-OS-4 cells overexpressed epidermal growth factor receptor , human epidermal growth factor receptor ( HER ) 2 , and phosphorylated HER2 protein .\n\nThe IC(50) values to cisplatin , paclitaxel , and lapatinib were 25.8 \\u03bcM , 686 nM , and 183 nM , respectively .\n\nHeterotransplantation in nude mice reflected the original tumor of the cells .\n\nThese results suggested that this cell line would be useful to study chemoresistant mechanisms and contribute to establishing novel treatment strategies for patients with ovarian cancer .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "N-Ras is one member of a family of oncoproteins that are commonly mutated in cancer .\n\nActivating mutations in N-Ras occur in a subset of colorectal cancers , but little in known about how the mutant protein contributes to onset and progression of the disease .\n\nUsing genetically engineered mice , we find that mutant N-Ras strongly promotes tumorigenesis in the context of inflammation .\n\nThe pro-tumorigenic nature of mutant N-Ras is related to its anti-apoptotic function , which is mediated by activation of a non-canonical MAPK pathway that signals through Stat3 .\n\nAs a result , inhibition of MEK selectively induces apoptosis in autochthonous colonic tumors expressing mutant N-Ras .\n\nThe translational significance of this finding is highlighted by our observation that NRAS mutation correlates with a less favorable clinical outcome for colorectal cancer patients .\n\nThese data demonstrate for the first time the important role that N-Ras plays in colorectal cancer .", "output": "Genomic instability and mutation, Resisting cell death" }, { "input": "Cells undergoing malignant transformation often exhibit a shift in cellular metabolism from oxidative phosphorylation to glycolysis .\n\nThis glycolytic shift , called the Warburg effect , provides a mechanistic basis for targeting glycolysis to suppress carcinogenesis through the use of dietary caloric restriction and energy restriction-mimetic agents ( ERMA ) .\n\nWe recently reported the development of a novel class of ERMAs that exhibits high potency in eliciting starvation-associated cellular responses and epigenetic changes in cancer cells though glucose uptake inhibition .\n\nThe lead ERMA in this class , OSU-CG5 , decreases the production of ATP and NADH in LNCaP prostate cancer cells .\n\nIn this study , we examined the effect of OSU-CG5 on the severity of preneoplastic lesions in male transgenic adenocarcinoma of the mouse prostate ( TRAMP ) mice .\n\nDaily oral treatment with OSU-CG5 at 100 mg/kg from 6 to 10 weeks of age resulted in a statistically significant decrease in the weight of urogenital tract and microdissected dorsal , lateral , and anterior prostatic lobes relative to vehicle controls .\n\nThe suppressive effect of OSU-CG5 was evidenced by marked decreases in Ki67 immunostaining and proliferating cell nuclear antigen ( PCNA ) expression in the prostate .\n\nOSU-CG5 treatment was not associated with evidence of systemic toxicity .\n\nMicroarray analysis indicated a central role for Akt , and Western blot analysis showed reduced phosphorylation and/or expression levels of Akt , Src , androgen receptor , and insulin-like growth factor-1 receptor in prostate lobes .\n\nThese findings support further investigation of OSU-CG5 as a potential chemopreventive agent .", "output": "Sustaining proliferative signaling" }, { "input": "It is well established that hyperplasia and decreased apoptosis of airway smooth muscle cells ( ASMCs ) play an important role in the asthmatic airway remodeling .\n\nTumor suppressor PTEN gene with phosphatase activity plays an important regulatory role in embryonic development , cell proliferation , and apoptosis , cell cycle regulation , migration ( invasion ) of the cytoskeleton .\n\nWe hypotheses that PTEN gene could affect the growth and viability of ASMCs through the regulation of PI3K/Akt , MAPK , and cell cycle-related gene expression .\n\nWe constructed a recombinant adenovirus to transfect ASMCs .\n\nCells were divided into the overexpression of PTEN gene group ( Ad-PTEN-GFP ) , negative control group ( Ad-GFP ) , and blank control group ( DMEM ) .\n\nThe cell apoptosis of ASMCs were evaluated by Hoechst-33342 staining and PE-7AAD double-labeled flow cytometry .\n\nThe cell cycle distribution was observed by flow cytometry with PI staining .\n\nThe expression of PTEN , p-Akt , total-Akt , p-ERK1/2 , total-ERK1/2 , cleaved-Caspases-3 , Caspases-9 , p21 , and Cyclin D1 were tested by the Western blotting .\n\nOur study revealed that overexpression of PTEN gene did not induce apoptosis of human ASMCs cultured in vitro .\n\nHowever , overexpression of PTEN inhibited proliferation of human ASMCs cultured in vitro and was associated with downregulation of Akt phosphorylation levels , while did not affect ERK1/2 phosphorylation levels .\n\nMoreover , overexpression of PTEN could induce ASMCs arrested in the G0/G1 phase through the downregulation of Cyclin D1 and upregulation of p21 expressions .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "Platinum compounds are the foundation of chemotherapy regimens for non-small cell lung cancer ( NSCLC ) despite poor response rates and limited response duration .\n\nIt has been reported that tumor expression of ERCC1 , a key component in nucleotide excision repair , may correlate with clinical response to platinum agents .\n\nWe found that most primary lung tumor specimens demonstrated a stronger protein expression of poly ( ADP ribose ) polymerases 1 ( PARP1 ) than their normal counterparts .\n\nWe therefore hypothesized that combining PARP inhibition with platinum compounds may be an approach to improve platinum-based therapy for NSCLC .\n\nDrug combination experiments revealed that two distinct PARP inhibitors , olaparib and veliparib not only potentiated the cell killing by cisplatin but also conferred cytotoxicity as a single agent specifically in ERCC1-low HCC827 and PC9 but not in ERCC1-high A549 and H157 lung cancer cells .\n\nMoreover , siRNA knockdown of ERCC1 in A549 and H157 cells increased their sensitivities to both cisplatin and olaparib in a synergistic manner in our model .\n\nFurthermore , mechanistic studies indicated that combined PARP inhibitor and cisplatin could lead to sustained DNA double strand breaks , prolonged G2/M cell cycle arrest with distinct activation of checkpoint kinase 1 signaling , and more pronounced apoptosis preferentially in lung cancer cells with low ERCC1 expression .\n\nCollectively , these data suggest that there is a synergistic relationship between PARP inhibition and low ERCC1 expression in NSCLC that could be exploited for novel therapeutic approaches in lung cancer therapy based on tumor ERCC1 expression .", "output": "Genomic instability and mutation, Evading growth suppressors, Resisting cell death" }, { "input": "In the present study , the question was addressed whether anthocyanins interfere with the topoisomerase I poison irinotecan in vivo .\n\nIn vivo complexes of enzyme to DNA bioassay was used to detect irinotecan-induced stabilization of topoisomerase I/DNA complexes and single cell gel electrophoresis to determine DNA-strand-break induction in the colon of male Wistar rats .\n\nFurthermore , analysis of anthocyanin concentrations in rat plasma and rat colon was included in the testing , demonstrating that anthocyanins reach the colon and the concentrations do not differ between rats that only received anthocyanins and the anthocyanin/irinotecan group .\n\nBlackberry extract was found to significantly reduce irinotecan-mediated topoisomerase I/DNA cleavable complex formation .\n\nOverall , anthocyanins did not notably increase cleavable complex formation .\n\nHowever , a significant increase of DNA damage was shown after a single dose of irinotecan as well as the single compounds cyanidin ( cy ) and cyanidin-3-glucoside ( cy-3-g ) .\n\nFurthermore , a significant reduction of irinotecan-induced DNA-strand breaks after a pretreatment with cy , cy-3-g and blackberry extract was observed .\n\nThus , the question arises whether anthocyanin-rich preparations might interfere with chemotherapy or whether , due to low systemic bioavailability , the preparations might provide protective potential in the gastrointestinal tract .", "output": "Genomic instability and mutation" }, { "input": "Peroxisome proliferator activated receptor \u03b3 ( PPAR\u03b3 ) , a subgroup of ligand-activated nuclear receptors , plays critical roles in cell cycle regulation , differentiation , apoptosis and invasion .\n\nPPAR\u03b3 is involved in tumorigenesis and is a potent target for cancer therapy .\n\nPPAR\u03b3 transactivation of KLF4 has been demonstrated in various studies ; however , how PPAR\u03b3 regulates KLF4 expression is not clear .\n\nIn this study , we revealed that PPAR\u03b3 regulates the expression of KLF4 by binding directly to the PPAR response element ( PPRE ) within the KLF4 promoter .\n\nThe PPRE resided at -1657bp to -1669bp upstream of the KLF4 ATG codon , which is essential for the transactivation of TGZ-induced KLF4 expression .\n\nFurthermore , we found that stable silencing of KLF4 obviously suppressed the G1/S arrest and anti-proliferation effects induced by PPAR\u03b3 ligands .\n\nTaken together , our data indicated that upregulating KLF4 upon PPAR\u03b3 activation is mediated through PPRE in KLF4 promoter , thus providing further insights into PPAR\u03b3 signal transduction pathway as well as a novel cancer therapeutic strategy .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "Splenic haemangiosarcomas ( HSAs ) from 122 dogs were characterized and classified according to their patterns of growth , survival time post splenectomy , metastases and chemotherapy .\n\nThe most common pattern of growth was a mixture of cavernous , capillary and solid tumour tissue .\n\nSurvival time post splenectomy was independent of the growth pattern ; however , it was influenced by chemotherapy and metastases .\n\nImmunohistochemical assessment of the expression of angiogenic factors ( fetal liver kinase-1 , angiopoietin-2 , angiopoietin receptor-2 and vascular endothelial growth factor A ) and conventional endothelial markers ( CD31 , factor VIII-related antigen ) revealed variable expression , particularly in undifferentiated HSAs .\n\nTherefore , a combination of endothelial markers should be used to confirm the endothelial origin of splenic tumours .", "output": "Inducing angiogenesis, Activating invasion and metastasis" }, { "input": "Bladder cancer ( BCa ) remained a major health problem .\n\nMed19 was related to tumor growth of BCa .\n\nBone morphogenetic proteins ( BMPs ) were reported to be critical in bone metastasis of cancer .\n\nWe therefore investigated the relations between Med19 and BMPs in BCa and their effect on bone metastasis of BCa .\n\nBladder cancer cell lines were cultured and interfered with Med19 shRNA and control .\n\nExpressions of BMP-1 , BMP-2 , BMP-4 , BMP-5 , BMP-6 , BMP-7 , BMP-9 , and BMP-15 were studied between 2 groups .\n\nFifty-two BCa samples were included for immunohistochemical staining of Med19 and BMP-2 .\n\nExpressions were scored and studied statistically .\n\nInvasiveness was studied with Transwell assay .\n\nSilencing or Med19 in BCa cells induced altered expressions of BMPs .\n\nIncreased expressions of BMP-1 , BMP-4 , BMP-6 , BMP-7 , and BMP-15 and decreased expressions of BMP-2 , BMP-5 , and BMP-9 were noticed , but only BMP-2 reached statistical significance .\n\nExpressions of Med19 and BMP-2 were significantly higher in cases with bone metastasis and were positively correlated in cases with bone metastasis and muscle invasion .\n\nMed19 is a critical factor involved in the invasiveness and promotion of bone metastasis of BCa , possibly via BMP-2 .", "output": "Activating invasion and metastasis" }, { "input": "Nectin-like molucule-5 ( Necl-5 ) is an immunoglobulin-like molecule that was originally identified as a poliovirus receptor and is often upregulated in cancer cells .\n\nIt has been said that Necl-5 plays a role in not only cell-cell adhesion , but also cell migration , proliferation , and metastasis .\n\nIn this study , we used a bronchioloalveolar carcinoma ( BAC ) cell line and fibroblasts to assess the expression of Necl-5 in the development of cancer-stroma communication by using an easy-to-prepare double-layered collagen gel hemisphere ( DL- CGH ) system that enables visualization of cell migration during invasion .\n\nThe expression of Necl-5 was higher in BAC cells than in fibroblasts .\n\nThis tendency didn't change even when the BAC cells were mixed with fibroblasts .\n\nTo assess the role of Necl-5 in the invasive activity of the BAC cells , we knocked down its expression using RNA interference ( RNAi ) .\n\nThe invasion assay with DL-CGH revealed that inhibitation of Necl-5 expression in the BAC cells was associated with suppressed invasiveness .\n\nIn addition , Necl-5 knockdown inhibited the movement and proliferation of the BAC cells .\n\nNecl-5 expression in lung cancer cells is crucial for their invasiveness in the cancer-stromal interaction , suggesting that Necl-5 could be a favorable molecular target for the suppression of invasiveness in lung adenocarcinoma .", "output": "Sustaining proliferative signaling, Activating invasion and metastasis" }, { "input": "AMP-activated protein kinase ( AMPK ) has been implicated in anti-proliferative actions in a range of cell systems .\n\nRecently , it was observed that Compound C , an inhibitor of AMPK , also reduced the cell viability in human diploid fibroblasts ( HDFs ) .\n\nCompound C-induced growth arrest was associated with a decrease in the cell cycle regulatory proteins , such as proliferating cell nuclear antigen , phosphorylated pRB , cyclin-dependent protein kinases ( Cdk 2 and 4 ) , cyclins ( D and E ) , and the Cdk inhibitors ( p21 , p16 , and p27 ) .\n\nTherefore , the present study examined the molecular mechanism of the antiproliferative effects of Compound C. Although Compound C inhibited serum-induced phosphorylation of Akt and its substrate , glycogen synthase kinase-3\u03b2 , it did not affect the Akt activity in vitro .\n\nCompound C significantly inhibited the receptor tyrosine phosphorylation and the activity of downstream signaling molecules , such as p85 phosphoinositide 3-kinase , phospholipase C-\u03b31 , and extracellular signal-regulated kinase 1/2 , induced by platelet-derived growth factor ( PDGF ) but not by epidermal growth factor- and insulin-like growth factor .\n\nIn vitro growth factor receptor tyrosine kinase activity profiling revealed the IC(50) for PDGF receptor-\u03b2 ( PDGFR\u03b2 ) to be 5.07 \u03bcM , whereas the IC(50) for the epidermal growth factor receptor and insulin-like growth factor receptor was \u2265100 \u03bcM .\n\nThe inhibitory effect of Compound C on PDGFR\u03b2 and Akt was also observed in AMPK\u03b1(1)/\u03b1(2)-knockout mouse embryonic fibroblasts , indicating that its inhibitory effect is independent of the AMPK activity .\n\nThe inhibitory effect of Compound C on cell proliferation and PDGFR\u03b2 tyrosine phosphorylation was also demonstrated in various PDGFR-expressing cells , including MRC-5 , BEAS-2B , rat aortic vascular smooth muscle cells , and A172 glioblastoma cells .\n\nThese results indicate that Compound C can be used as a potential antiproliferative agent for PDGF- or PDGFR-associated diseases , such as cancer , atherosclerosis , and fibrosis .", "output": "Sustaining proliferative signaling" }, { "input": "SMG1 is a member of the phosphoinositide kinase-like kinase family of proteins that includes ATM , ATR , and DNA-PK , proteins with known roles in DNA damage and cellular stress responses .\n\nSMG1 has a well-characterized role in nonsense-mediated decay as well as suggested roles in the DNA damage response , resistance to oxidative stress , regulation of hypoxic responses , and apoptosis .\n\nTo understand the roles of SMG1 further , we generated a Genetrap Smg1 mouse model .\n\nSmg1 homozygous KO mice were early embryonic lethal , but Smg1 heterozygous mice showed a predisposition to a range of cancers , particularly lung and hematopoietic malignancies , as well as development of chronic inflammation .\n\nThese mice did not display deficiencies in known roles of SMG1 , including nonsense-mediated decay .\n\nHowever , they showed elevated basal tissue and serum cytokine levels , indicating low-level inflammation before the development of tumors .\n\nSmg1 heterozygous mice also showed evidence of oxidative damage in tissues .\n\nThese data suggest that the inflammation observed in Smg1 haploinsufficiency contributes to susceptibility to cancer and that Smg1-deficient animals represent a model of inflammation-enhanced cancer development .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "BACKGROUND O(6) -methylguanine-DNA methyltransferase ( MGMT ) is a DNA repair enzyme that can protect cells from carcinogenic effects of alkylating agents by removing adducts from the O(6) position of guanine .\n\nEvidences indicated that areca quid chewing may increase the risk of oral squamous cell carcinoma ( OSCC ) .\n\nThis study was to investigate the role of MGMT expression in OSCCs and the normal oral tissues .\n\nMETHODS Thirty-two OSCCs from areca quid chewers and ten normal oral tissue biopsy samples without areca quid chewing were analyzed by the immunohistochemistry for MGMT .\n\nPrimary human oral keratinocytes ( HOKs ) were challenged with arecoline , the major alkaloid of areca nut , by Western blot .\n\nNicotine , an important component of cigarette smoke , was added to find the possible regulatory mechanisms .\n\nRESULTS Significant association was observed between low MGMT expression and advanced clinical stage of OSCCs and lymph node metastasis ( P=0.03 ) .\n\nMGMT expression was significantly higher in patients only chewing areca quid than patients both chewing areca quid and smoking ( P=0.028 ) .\n\nArecoline was found to elevate MGMT expression in a dose- and time-dependent manner .\n\nThe addition of nicotine was found to enhance arecoline-induced MGMT expression .\n\nCONCLUSION Our results indicate that MGMT could be used clinically as a predictive marker for tumor processing , the potential for lymph node metastasis as well as advanced clinical stage .\n\nMGMT expression was significantly upregulated by arecoline in HOKs .\n\nNicotine has a synergistic effect of arecoline-induced MGMT expression .\n\nThe cigarette smoking may act synergistically in the pathogenesis of OSCC in areca quid chewers via the upregulation of MGMT .", "output": "Genomic instability and mutation, Activating invasion and metastasis" }, { "input": "The ALKBH family of proteins are highly expressed in various types of human cancer where they are involved in tumor growth and progression .\n\nHowever , multiple isoforms of ALKBH exist and the effect of individual isoforms on the development of urinary bladder cancer is unknown , particularly the molecular mechanisms involved in the progression from a noninvasive to invasive phenotype .\n\nWe examined the role and function of ALKBH2 in human bladder cancer development in vitro and provide the first report that suppression of ALKBH2 in a human urothelial carcinoma cell line , KU7 , reduced the expression of the transmembrane mucin protein , MUC1 , and induced G1 cell cycle arrest .\n\nMoreover , reduction of ALKBH2 suppressed epithelial to mesenchymal transition ( EMT ) via increasing E-cadherin and decreasing vimentin expression .\n\nTransfection of MUC1 siRNA inhibited cell proliferation and EMT to the same extent as ALKBH2 gene silencing in vitro .\n\nALKBH2 knockdown significantly suppressed MUC1 expression and tumor volume of bladder cancers in vivo as assessed in an orthotopic mouse model using ALKBH2 shRNA transfected KU7 cells .\n\nImmunohistochemical examination showed high expression levels of ALKBH2 in human urothelial carcinoma samples , especially in high-grade , superficially and deeply invasive carcinomas ( pT(1) and >pT(2) ) , and in carcinoma in situ but not in normal urothelium .\n\nThis study demonstrates that ALKBH2 is an upstream molecule of the oncoprotein , MUC1 , and regulates cell cycle and EMT , resulting in progression of urothelial carcinomas .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Activating invasion and metastasis" }, { "input": "Sonodynamic therapy ( SDT ) has shown great potential in target cancer therapy , but it induced cell death modes hasn't been fully investigated .\n\nThis study was to examine autophagic and apoptotic responses to protoporphyrin IX ( PpIX ) mediated SDT in murine leukemia L1210 cells .\n\nAfter SDT , the occurrence of autophagy was identified by morphological observation and biochemical analysis .\n\nMeanwhile , the mitochondria dependent apoptosis pathway was examined to participate in SDT induced cell death .\n\nThe relationship between autophagy and apoptosis was further investigated by applying pharmacological inhibition studies , which indicated that impairment of autophagy enhanced the anti-tumor effect of SDT through induction of apoptosis and necrosis , while caspase inhibition didn't affect autophagic vacuoles formation or protect SDT induced cytotoxicity .\n\nThe findings supported that autophagic vacuoles formed upstream and independently from caspase-dependent cell death .\n\nAdditionally , the possible mechanism of SDT-induced autophagy was evaluated by measurement of ROS ( reactive oxygen species ) formation .\n\nResult suggested ROS play important role in initiating autophagy , possibly through the sono-damaged mitochondria being enclosed by autophagic vacuoles .\n\nAll together , these data indicate that autophagy may be cytoprotective in our experimental system , and point to an important insight into how autophagy inhibitors , in combination with SDT may contribute a regimen for cancer therapy .", "output": "Tumor promoting inflammation, Resisting cell death" }, { "input": "The Wnt/\u03b2-catenin pathway regulates the viability and radiosensitivity of head and neck squamous cancer cells ( HNSCC ) .\n\nIncreased \u03b2-catenin predisposes HNSCC patients to poor prognosis and survival .\n\nThis study was conducted to determine the mechanism by which \u03b2-catenin regulates the viability of HNSCC .\n\nAMC-HN-3 , -HN-8 , UM-SCC-38 , and -SCC-47 cells , which were established from human head and neck cancer specimens , underwent cell death following \u03b2-catenin silencing. \u03b2-Catenin silencing significantly induced G1 arrest and increased the expression of Bax and active caspase-3 , which demonstrates the sequential activation of apoptotic cascades following treatment of HNSCC with targeted siRNA .\n\nIntriguingly , \u03b2-catenin silencing also induced autophagy .\n\nHere , we confirm that the number of autophagic vacuoles and the expression of type II light chain 3 were increased in cells that were treated with \u03b2-catenin siRNA .\n\nThese cell death modes are most likely due to the activation of LKB1-dependent AMPK following \u03b2-catenin silencing .\n\nThe activated LKB1/AMPK pathway in AMC-HN-3 cells caused G1 arrest by phosphorylating p53 and suppressing mTOR signaling .\n\nIn addition , treating AMC-HN-3 cells with LKB1 siRNA preserved cell viability against \u03b2-catenin silencing-induced cytotoxicity .\n\nTaken together , these results imply that following \u03b2-catenin silencing , HNSCC undergo both apoptotic and autophagic cell death that are under the control of LKB1/AMPK .\n\nTo the best of our knowledge , these results suggest for the first time that novel crosstalk between \u03b2-catenin and the LKB1/AMPK pathway regulates the viability of HNSCC .\n\nThis study thus presents new insights into our understanding of the cellular and molecular mechanisms involved in \u03b2-catenin silencing-induced cell death .", "output": "Resisting cell death" }, { "input": "Colorectal cancer ( CRC ) is one of the leading causes of cancer deaths in Western countries .\n\nA significant number of CRC patients undergoing curatively intended surgery subsequently develop recurrence and die from the disease .\n\nMicroRNAs ( miRNAs ) are aberrantly expressed in cancers and appear to have both diagnostic and prognostic significance .\n\nIn this study , we identified novel miRNAs associated with recurrence of CRC , and their possible mechanism of action .\n\nTaqMan\ufffd Human MicroRNA Array Set v2.0 was used to profile the expression of 667 miRNAs in 14 normal colon mucosas and 46 microsatellite stable CRC tumors .\n\nFour miRNAs ( miR-362-3p , miR-570 , miR-148a* and miR-944 ) were expressed at a higher level in tumors from patients with no recurrence ( p < 0.015 ) , compared to tumors from patients with recurrence .\n\nA significant association with increased disease free survival was confirmed for miR-362-3p in a second independent cohort of 43 CRC patients , using single TaqMan\ufffd microRNA assays .\n\nIn vitro functional analysis showed that over-expression of miR-362-3p in colon cancer cell lines reduced cell viability , and proliferation mainly due to cell cycle arrest .\n\nE2F1 , USF2 and PTPN1 were identified as potential miR-362-3p targets by mRNA profiling of HCT116 cells over-expressing miR-362-3p .\n\nSubsequently , these genes were confirmed as direct targets by Luciferase reporter assays .\n\nTheir knockdown in vitro phenocopied the effects of miR-362-3p over-expression .\n\nWe conclude that miR-362-3p may be a novel prognostic marker in CRC , and hypothesize that the positive effects of augmented miR-362-3p expression may in part be mediated through the targets E2F1 , USF2 and PTPN1. \ufffd 2012 Wiley Periodicals , Inc .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "Dichlorvos , an organophosphate ( OP ) , is known to cause oxidative stress in the central nervous system ( CNS ) .\n\nPreviously we have shown that dichlorvos treatment promoted the levels of proinflammatory molecules and ultimately induced apoptotic cell death in primary microglial cells .\n\nHere we studied the effect of dichlorvos on crucial cell cycle regulatory proteins and the DNA damage sensor ataxia-telangiectasia mutated ( ATM ) .\n\nWe found a significant increase in p53 and its downstream target , p21 , levels in dichlorvos-treated microglial cells compared with control cells .\n\nMoreover , dichlorvos exposure promoted the levels of different cell cycle regulatory proteins .\n\nThese results along with flow cytometry results suggested that primary microglial cells were arrested at G1 and G2/M phase after dichlorvos exposure .\n\nWe have shown in a previous study that dichlorvos can induce DNA damage in microglia ; here we found that microglial cells also tried to repair this damage by inducing a DNA repair enzyme , i.e. , ATM .\n\nWe observed a significant increase in the levels of ATM after dichlorvos treatment compared with control .", "output": "Evading growth suppressors" }, { "input": "Growth differentiation factor-15 ( GDF-15 ) and the CCN family member , connective tissue growth factor ( CCN2 ) , are associated with cardiac disease , inflammation and cancer .\n\nThe precise role and signaling mechanism for these factors in normal and diseased tissues remains elusive .\n\nHere we demonstrate an interaction between GDF-15 and CCN2 using yeast two-hybrid assays and have mapped the domain of interaction to the von Willebrand factor type C domain of CCN2 .\n\nBiochemical pull down assays using secreted GDF-15 and His-tagged CCN2 produced in PC-3 prostate cancer cells confirmed a direct interaction between these proteins .\n\nTo investigate the functional consequences of this interaction , in vitro angiogenesis assays were performed .\n\nWe demonstrate that GDF-15 blocks CCN2-mediated tube formation in human umbilical vein endothelial ( HUVEC ) cells .\n\nTo examine the molecular mechanism whereby GDF-15 inhibits CCN2-mediated angiogenesis , activation of \u03b1(V) \u03b2(3) integrins and focal adhesion kinase ( FAK ) was examined .\n\nCCN2-mediated FAK activation was inhibited by GDF-15 and was accompanied by a decrease in \u03b1(V) \u03b2(3) integrin clustering in HUVEC cells .\n\nThese results demonstrate , for the first time , a novel signaling pathway for GDF-15 through interaction with the matricellular signaling molecule CCN2 .\n\nFurthermore , antagonism of CCN2 mediated angiogenesis by GDF-15 may provide insight into the functional role of GDF-15 in disease states .\n\nJ. Cell .\n\nBiochem. \ufffd 2012 Wiley Periodicals , Inc .", "output": "Inducing angiogenesis" }, { "input": "MD-Fraction is a highly purified soluble \u03b2-glucan derived from Grifola frondosa ( an oriental edible mushroom ) .\n\nIntraperitoneal ( i.p. ) injection of MD-Fraction has been reported to inhibit tumor growth via enhancement of the host immune system .\n\nIn this study , we demonstrated that oral administration of MD-Fraction as well as i.p. injection significantly inhibited tumor growth in murine tumor models .\n\nAfter oral administration , MD-Fraction was not transferred to the blood in its free form but was captured by antigen-presenting cells such as macrophages and dendritic cells ( DCs ) present in the Peyer's patch .\n\nThe captured MD-Fraction was then transported to the spleen , thereby inducing the systemic immune response .\n\nOur study showed that MD-Fraction directly induced DC maturation via a C-type lectin receptor dectin-1 pathway .\n\nThe therapeutic response of orally administered MD-Fraction was associated with ( i ) induced systemic tumor-antigen specific T cell response via dectin-1-dependent activation of DCs , ( ii ) increased infiltration of the activated T cells into the tumor , and ( iii ) decreased number of tumor-caused immunosuppressive cells such as regulatory T cells and myeloid-derived suppressor cells .\n\nOur preclinical study suggests that MD-Fraction is a useful oral therapeutic agent in the management of patients with cancer. \ufffd 2012 Wiley Periodicals , Inc .", "output": "Avoiding immune destruction" }, { "input": "Aurora kinases play an essential role in mitotic progression and are potentially druggable targets in cancer therapy .\n\nWe identified benzo[e]pyridoindoles ( BePI ) as powerful aurora kinase inhibitors .\n\nTheir efficiency was demonstrated both in enzymatic inhibition studies and in cell culture assays .\n\nNew BePI molecules were synthesized , and a structure-activity relationship study was conducted with the aim of improving the activity and solubility of the lead compound .\n\nTetracyclic BePI derivatives are characterized by a particular curved shape , and the presence of an oxo group on the pyridine ring was found to be required for aurora kinase\u2005B inhibition .\n\nNew hydrosoluble benzo[e]pyridoindolones were subsequently designed , and their efficacy was tested by a combination of cell-cycle analysis and time-lapse experiments in live cells .\n\nThe most active BePI derivative , 13\u2009b , inhibited the cell cycle , drove cells to polyploidy , and eventually induced apoptosis .\n\nIt exhibited high antiproliferative activity in HeLa cells with an IC(50) value of 63\u2005nM .\n\nRelative to compounds tested in clinical trials , this antiproliferative potency places 13\u2009b among the top 10 aurora kinase inhibitors .\n\nOur results justify further in\u2005vivo evaluation in preclinical animal models of cancer .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "Elevated androgen receptor ( AR ) activity in castration-resistant prostate cancer ( CRPC ) may occur through increased levels of AR coactivator proteins .\n\nVav3 , a guanine nucleotide exchange factor ( GEF ) , is upregulated following progression to castration-resistance in preclinical models and is overexpressed in a significant number of human prostate cancers .\n\nVav3 is a novel coactivator of the AR .\n\nWe sought to identify Vav3 binding partners in an effort to understand the molecular mechanisms underlying Vav3 enhancement of AR activity and to identify new therapeutic targets .\n\nThe cell division cycle 37 homolog ( Cdc37 ) , a protein kinase-specific co-chaperone for Hsp90 , was identified as a Vav3 interacting protein by yeast two hybrid screening .\n\nVav3-Cdc37 interaction was confirmed by GST pulldown and , for native proteins , by coimmunoprecipitation experiments in prostate cancer cells .\n\nCdc37 potentiated Vav3 coactivation of AR transcriptional activity and Vav3 enhancement of AR amino-carboxyl terminal ( N-C ) interaction , which is essential for optimal receptor transcriptional activity .\n\nCdc37 increased prostate cancer cell proliferation selectively in Vav3 expressing cells .\n\nCdc37 did not affect Vav3 nucleotide exchange activity , Vav3 protein levels or subcellular localization .\n\nDisruption of Vav3-Cdc37 interaction inhibited Vav3 enhancement of AR transcriptional activity and AR N-C interaction .\n\nDiminished Vav3-Cdc37 interaction also caused decreased prostate cancer cell proliferation selectively in Vav3 expressing cells .\n\nTaken together , we identified a novel Vav3 interacting protein that enhances Vav3 coactivation of AR and prostate cancer cell proliferation .\n\nVav3-Cdc37 interaction may provide a new therapeutic target in prostate cancer .", "output": "Sustaining proliferative signaling" }, { "input": "The anti-tumor antibiotic salinomycin ( Sal ) was recently identified as a selective inhibitor of breast cancer stem cells ; however , the effect of Sal on hepatocellular carcinoma ( HCC ) is not clear .\n\nThis study aimed to determine the anti-tumor efficacy and mechanism of Sal on HCC .\n\nHCC cell lines ( HepG2 , SMMC-7721 , and BEL-7402 ) were treated with Sal .\n\nCell doubling time was determinated by drawing growth curve , cell viability was evaluated using the Cell Counting Kit 8 .\n\nThe fraction of CD133(+) cell subpopulations was assessed by flow cytometry .\n\nWe found that Sal inhibits proliferation and decreases PCNA levels as well as the proportion of HCC CD133(+)cell subpopulations in HCC cells .\n\nCell cycle was analyzed using flow cytometry and showed that Sal caused cell cycle arrest of the various HCC cell lines in different phases .\n\nCell apoptosis was evaluated using flow cytometry and Hoechst 33342 staining .\n\nSal induced apoptosis as characterized by an increase in the Bax/Bcl-2 ratio .\n\nSeveral signaling pathways were selected for further mechanistic analyses using real time-PCR and Western blot assays .\n\nCompared to control , \u03b2-catenin expression is significantly down-regulated upon Sal addition .\n\nThe Ca(2+) concentration in HCC cells was examined by flow cytometry and higher Ca(2+) concentrations were observed in Sal treatment groups .\n\nThe anti-tumor effect of Sal was further verified in vivo using the hepatoma orthotopic tumor model and the data obtained showed that the size of liver tumors in Sal-treated groups decreased compared to controls .\n\nImmunohistochemistry and TUNEL staining also demonstrated that Sal inhibits proliferation and induces apoptosis in vivo .\n\nFinally , the role of Sal on in vivo Wnt/\u03b2-catenin signaling was evaluated by Western blot and immunohistochemistry .\n\nThis study demonstrates Sal inhibits proliferation and induces apoptosis of HCC cells in vitro and in vivo and one potential mechanism is inhibition of Wnt/\u03b2-catenin signaling via increased intracellular Ca(2+) levels .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "The pancreatic stellate cells ( PSCs ) have complex roles in pancreas , including tissue repair and fibrosis .\n\nPSCs surround ATP releasing exocrine cells , but little is known about purinergic receptors and their function in PSCs .\n\nOur aim was to resolve whether PSCs express the multifunctional P2X7 receptor and elucidate how it regulates PSC viability .\n\nThe number of PSCs isolated from wild type ( WT ) mice was 50% higher than those from the Pfizer P2X7 receptor knock out ( KO ) mice .\n\nThe P2X7 receptor protein and mRNA of all known isoforms were expressed in WT PSCs , while KO PSCs only expressed truncated versions of the receptor .\n\nIn culture , the proliferation rate of the KO PSCs was significantly lower .\n\nInclusion of apyrase reduced the proliferation rate in both WT and KO PSCs , indicating importance of endogenous ATP .\n\nExogenous ATP had a two-sided effect .\n\nProliferation of both WT and KO cells was stimulated with ATP in a concentration-dependent manner with a maximum effect at 100 \u00b5M .\n\nAt high ATP concentration ( 5 mM ) , WT PSCs , but not the KO PSCs died .\n\nThe intracellular Ca(2+) signals and proliferation rate induced by micromolar ATP concentrations were inhibited by the allosteric P2X7 receptor inhibitor az10606120 .\n\nThe P2X7 receptor-pore inhibitor A438079 partially prevented cell death induced by millimolar ATP concentrations .\n\nThis study shows that ATP and P2X7 receptors are important regulators of PSC proliferation and death , and therefore might be potential targets for treatments of pancreatic fibrosis and cancer .", "output": "Sustaining proliferative signaling" }, { "input": "CDA-2 ( cell differentiation agent 2 ) , a urinary preparation , has potent anti- proliferative and pro-apoptotic properties in cancer cells .\n\nHowever , the mechanisms of tumor inhibitory action of CDA-2 are far from clear , and especially there was no report on lung cancer .\n\nHere we demonstrate that CDA-2 and its main component phenylacetylglutamine ( PG ) reduce the metastatic lung tumor growth , and increases survival time after inoculation with Lewis lung carcinoma ( LLC ) cells in a dose-dependent manner in C57BL6 mice .\n\nProliferative program analysis in cancer cells revealed a fundamental impact of CDA-2 and PG on proliferation and apoptosis , including Bcl-2 , Bcl-XL , cIAP1 , Survivin , PCNA , Ki-67 proteins and TUNEL assays .\n\nCDA-2 and PG significantly reduced NF-\u03baB DNA-binding activity in lung cancer cells and in alveolar macrophages of tumor bearing mice and especially decreased the release of inflammatory factors including TNF\u03b1 , IL-6 , and KC .\n\nFurthermore , CDA-2 and PG decrease the expressions of TLR2 , TLR6 , and CD14 , but not TLR1 , TLR3 , TLR4 , and TLR9 in bone-marrow-derived macrophages ( BMDM ) of mice stimulated by LLC-conditioned medium ( LLC-CM ) .\n\nOver-expressing TLR2 in BMDM prevented CDA-2 and PG from inhibiting NF-\u03baB activation , as well as induction of TNF\u03b1 and IL-6 .\n\nTLR2:TLR6 complexes mediate the effect of NF-\u03baB inactivation by CDA-2 .\n\nIn conclusion , CDA-2 potently inhibits lung tumor development by reduction of the inflammation in lung through suppression of NF-\u03baB activation in myeloid cells , associating with modulation of TLR2 signaling .", "output": "Tumor promoting inflammation, Resisting cell death, Activating invasion and metastasis" }, { "input": "Hepatocellular carcinoma ( HCC ) is the most prevalent liver tumor and a deadly disease with limited therapeutic options .\n\nDysregulation of cell signaling pathways is a common denominator in tumorigenesis , including hepatocarcinogenesis .\n\nThe epidermal growth factor receptor ( EGFR ) signaling system is commonly activated in HCC , and is currently being evaluated as a therapeutic target in combination therapies .\n\nWe and others have identified a central role for the EGFR ligand amphiregulin ( AR ) in the proliferation , survival and drug resistance of HCC cells .\n\nAR expression is frequently up-regulated in HCC tissues and cells through mechanisms not completely known .\n\nHere we identify the \u03b2-catenin signaling pathway as a novel mechanism leading to transcriptional activation of the AR gene in human HCC cells .\n\nActivation of \u03b2-catenin signaling , or expression of the T41A \u03b2-catenin active mutant , led to the induction of AR expression involving three specific \u03b2-catenin-Tcf responsive elements in its proximal promoter .\n\nWe demonstrate that HCC cells expressing the T41A \u03b2-catenin active mutant show enhanced proliferation that is dependent in part on AR expression and EGFR signaling .\n\nWe also demonstrate here a novel cross-talk of the EGFR system with fibroblast growth factor 19 ( FGF19 ) .\n\nFGF19 is a recently identified driver gene in hepatocarcinogenesis and an activator of \u03b2-catenin signaling in HCC and colon cancer cells .\n\nWe show that FGF19 induced AR gene expression through the \u03b2-catenin pathway in human HCC cells .\n\nImportantly , AR up-regulation and EGFR signaling participated in the induction of cyclin D1 and cell proliferation elicited by FGF19 .\n\nFinally , we demonstrate a positive correlation between FGF19 and AR expression in human HCC tissues , therefore supporting in clinical samples our experimental observations .\n\nThese findings identify the AR/EGFR system as a key mediator of FGF19 responses in HCC cells involving \u03b2-catenin signaling , and suggest that combined targeting of FGF19 and AR/EGFR may enhance therapeutic efficacy .", "output": "Sustaining proliferative signaling" }, { "input": "Lung cancers express the cholinergic autocrine loop , which facilitates the progression of cancer cells .\n\nThe antagonists of mAChRs have been demonstrated to depress the growth of small cell lung cancers ( SCLCs ) .\n\nIn this study we intended to investigate the growth inhibitory effect of R2HBJJ , a novel muscarinic antagonist , on non-small cell lung cancer ( NSCLC ) cells and the possible mechanisms .\n\nThe competitive binding assay revealed that R2HBJJ had a high affinity to M3 and M1 AChRs .\n\nR2HBJJ presented a strong anticholinergic activity on carbachol-induced contraction of guinea-pig trachea .\n\nR2HBJJ markedly suppressed the growth of NSCLC cells , such as H1299 , H460 and H157 .\n\nIn H1299 cells , both R2HBJJ and its leading compound R2-PHC displayed significant anti-proliferative activity as M3 receptor antagonist darifenacin .\n\nExogenous replenish of ACh could attenuate R2HBJJ-induced growth inhibition .\n\nSilencing M3 receptor or ChAT by specific-siRNAs resulted in a growth inhibition of 55.5% and 37.9% on H1299 cells 96 h post transfection , respectively .\n\nFurther studies revealed that treatment with R2HBJJ arrested the cell cycle in G0/G1 by down-regulation of cyclin D1-CDK4/6-Rb .\n\nTherefore , the current study reveals that NSCLC cells express an autocrine and paracrine cholinergic system which stimulates the growth of NSCLC cells .\n\nR2HBJJ , as a novel mAChRs antagonist , can block the local cholinergic loop by antagonizing predominantly M3 receptors and inhibit NSCLC cell growth , which suggest that M3 receptor antagonist might be a potential chemotherapeutic regimen for NSCLC .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "DNA repair is an essential cellular process required to maintain genomic stability .\n\nEvery cell is subjected to thousands of DNA lesions daily under normal physiological conditions .\n\nIonizing radiation ( IR ) is a major DNA damaging agent that can be produced by both natural and man-made sources .\n\nA common source of radiation exposure is through its use in medical diagnostics or treatments such as for cancer radiotherapy where relatively high doses are received by patients .\n\nTo understand the detailed DNA repair gene transcription response to high dose IR , gene expression exon array studies have been performed and the response to radiation in two divergent cell types , lymphoblastoid cell lines and primary fibroblasts , has been examined .\n\nThese exon arrays detect expression levels across the entire gene , and have the advantage of high sensitivity and the ability to identify alternative transcripts .\n\nWe found a selection of DNA repair genes , including some not previously reported , that are modulated in response to radiation .\n\nDetailed dose and time course kinetics of DNA repair transcription was conducted and results have been validated utilizing PCR methods .\n\nAlternative transcription products in response to IR were identified in several DNA repair genes including RRM2B and XPC where alternative initiation sites were found .\n\nThese investigations have advanced the knowledge about the transcriptional response of DNA repair .", "output": "Genomic instability and mutation" }, { "input": "Prostate cancer is the second most common cancer in elderly men worldwide and its incidence rate is rising continuously .\n\nAgents capable of inducing apoptosis in prostate cancer cells seem a promising approach to treat this malignancy .\n\nIn this study we describe the synthesis of a number of novel N- and N,N'-substituted S-2,3,4,5,6-pentabromobenzylisothiouronium bromides and their activity against the human prostate adenocarcinoma PC3 cell line .\n\nAll the compounds produced changes in mitochondrial transmembrane potential and cell cycle progression , showed a cytostatic effect and induced apoptosis in the tested cancer line in a concentration- and time-dependent manner .\n\nThe most effective compounds ZKK-3 , ZKK-9 and ZKK-13 produced , at 20 microM concentration , apoptosis in 42 , 46 , and 66% of the cells , respectively , after 48 h incubation .\n\nTwo selected S-2,3,4,5,6-pentabromobenzylisothiouronium bromides ( ZKK-3 , ZKK-9 ) showed also a synergic proapoptotic effect with the new casein kinase II inhibitor 2-(4-methylpiperazin-1-yl)-4,5,6,7-tetrabromo-1H-benzimidazole ( TBIPIP ) in the PC3 cell line .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "Objective To investigate the effect of bufalin on nucleus-mitochondria localization of human telomerase reverse transcriptase(hTERT) by exploring its effect on proliferation and apoptosis in human esophageal squamous carcinoma EC9706 cells .\n\nMethods EC9706 cells were treated with bufalin at various concentrations , and then the cell growth inhibition of EC9706 cells was examined by CCK-8 assay and the 50% inhibitory concentration ( IC(50) ) was calculated.Cell cycle analysis was performed by flow cytometry with PI staining , and nucleus morphology of apoptosis were observed by fluorescence microscopy with Hoechst 33342 staining .\n\nThe apoptotic index was measured by flow cytometry with Annexin V-FITC/PI double staining. hTERT subcellular localization and protein expression were determined by Western blotting and multiple immunofluorescence labling combined with laser confocal scanning microscopy .\n\nResults The proliferation of EC 9706 cells was significantly inhibited by bufalin along with the increase of processing time and concentrations ( p<0.01 ) .\n\nAfter the EC9706 cells were exposed to 100 nmol/L bufalin,the number of cells gradually decreased in G(1) phase and increased in S and G(2)/M phases(p<0.05) .\n\nThe typical nucleus morphological changes of apoptosis were observed and the apoptotic index was increased(p<0.01) .\n\nThe expression of hTERT decreased in nucleus but increased in mitochondria(p<0.05) .\n\nConclusions Bufalin can inhibit the proliferation of human esophageal squamous carcinoma EC9706 cells in a time- and dose-dependent manner .\n\nIt can arrest cell cycle in S and G(2)/M phases and induce the apoptosis of EC 9706 cells. hTERT is localized in both nucleus and mitochondria,and can be partially translocated from nucleus to mitochondria during the bufalin-induced apoptosis .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "Induced pluripotent stem ( iPS ) cells share some basic properties , such as self-renewal and pluripotency , with cancer cells , and they also appear to share several metabolic alterations that are commonly observed in human tumors .\n\nThe cancer cells ' glycolytic phenotype , first reported by Otto Warburg , is necessary for the optimal routing of somatic cells to pluripotency .\n\nHowever , how iPS cells establish a Warburg-like metabolic phenotype and whether the metabolic pathways that support the bioenergetics of iPS cells are produced by the same mechanisms that are selected during the tumorigenic process remain largely unexplored .\n\nWe recently investigated whether the reprogramming-competent metabotype of iPS cells involves changes in the activation/expression status of the H ( + ) -ATPase , which is a core component of mitochondrial oxidative phosphorylation that is repressed at both the activity and protein levels in human carcinomas , and of the lipogenic switch , which refers to a marked overexpression and hyperactivity of the acetyl-CoA carboxylase ( ACACA ) and fatty acid synthase ( FASN ) lipogenic enzymes that has been observed in nearly all examined cancer types .\n\nA comparison of a starting population of mouse embryonic fibroblasts and their iPS cell progeny revealed that somatic cell reprogramming involves a significant increase in the expression of ATPase inhibitor factor 1 ( IF1 ) , accompanied by extremely low expression levels of the catalytic \u03b2-F1-ATPase subunit .\n\nThe pharmacological inhibition of ACACA and FASN activities markedly decreases reprogramming efficiency , and ACACA and FASN expression are notably upregulated in iPS cells .\n\nImportantly , iPS cells exhibited a significant intracellular accumulation of neutral lipid bodies ; however , these bodies may be a reflection of intense lysosomal/autophagocytic activity rather than bona fide lipid droplet formation in iPS cells , as they were largely unresponsive to pharmacological modulation of PPARgamma and FASN activities .\n\nThe AMPK agonist metformin , which endows somatic cells with a bioenergetic infrastructure that is protected against reprogramming , was found to drastically elongate fibroblast mitochondria , fully reverse the high IF1/\u03b2-F1-ATPase ratio and downregulate the ACACA/FASN lipogenic enzymes in iPS cells .\n\nThe mitochondrial H ( + ) -ATP synthase and the ACACA/FASN-driven lipogenic switch are newly characterized as instrumental metabolic events that , by coupling the Warburg effect to anabolic metabolism , enable de-differentiation during the reprogramming of somatic cells to iPS cells .", "output": "Cellular energetics" }, { "input": "Recent studies have highlighted an apparently paradoxical link between self-renewal and senescence triggered by DNA damage in certain cell types .\n\nIn addition , the finding that TP53 can suppress senescence has caused a re-evaluation of its functional role in regulating these outcomes .\n\nTo investigate these phenomena and their relationship to pluripotency and senescence , we examined the response of the TP53-competent embryonal carcinoma ( EC ) cell line PA1 to etoposide-induced DNA damage .\n\nNuclear POU5F1/OCT4A and p21cip1 were upregulated in the same cells following etoposide-induced G 2M arrest .\n\nHowever , while accumulating in the karyosol , the amount of OCT4A was reduced in the chromatin fraction .\n\nPhosphorylated CHK2 and Rad51/\u03b3H2AX-positive nuclear foci , overexpression of Aurora B kinase and moderate macroautophagy were evident .\n\nUpon release from G 2M arrest , cells with repaired DNA entered mitoses , while the cells with persisting DNA damage remained at this checkpoint or underwent mitotic slippage and gradually senesced .\n\nReduction of TP53 using sh- or si-RNA prevented the upregulation of OCT4A and p21cip1 and increased DNA damage .\n\nSubsequently , mitoses , micronucleation and senescence were all enhanced after TP53 reduction with senescence confirmed by upregulation of CDKN2A/p16ink4a and increased sa-\u03b2-galactosidase positivity .\n\nThose mitoses enhanced by TP53 silencing were shown to be multicentrosomal and multi-polar , containing fragmented and highly deranged chromosomes , indicating a loss of genome integrity .\n\nTogether , these data suggest that TP53-dependent coupling of self-renewal and senescence pathways through the DNA damage checkpoint provides a mechanism for how embryonal stem cell-like EC cells safeguard DNA integrity , genome stability and ultimately the fidelity of self-renewal .", "output": "Genomic instability and mutation, Enabling replicative immortality, Evading growth suppressors" }, { "input": "Filamin-A cross-links actin filaments into dynamic orthogonal networks , and interacts with an array of proteins of diverse cellular functions .\n\nBecause several filamin-A interaction partners are implicated in signaling of cell mobility regulation , we tested the hypothesis that filamin-A plays a role in cancer metastasis .\n\nUsing four pairs of filamin-A proficient and deficient isogenic cell lines , we found that filamin-A deficiency in cancer cells significantly reduces their migration and invasion .\n\nUsing a xenograft tumor model with subcutaneous and intracardiac injections of tumor cells , we found that the filamin-A deficiency causes significant reduction of lung , splenic and systemic metastasis in nude mice .\n\nWe evaluated the expression of filamin-A in breast cancer tissues by immunohistochemical staining , and found that low levels of filamin-A expression in cancer cells of the tumor tissues are associated with a better distant metastasis-free survival than those with normal levels of filamin-A .\n\nThese data not only validate filamin-A as a prognostic marker for cancer metastasis , but also suggest that inhibition of filamin-A in cancer cells may reduce metastasis and that filamin-A can be used as a therapeutic target for filamin-A positive cancer .", "output": "Activating invasion and metastasis" }, { "input": "The increasing evidence supported the role of Enhancer of Zeste Homolog 2 ( EZH2 ) in the cancer development and progression .\n\nHowever , its precise role in the tumorigenesis of Nasopharyngeal Carcinoma ( NPC ) remains to be elucidated .\n\nEZH2 was depleted by retroviral infection in the NPC cells ( HK-1 , CNE-2 , CNE-1 and C666-1 ) .\n\nThe degree of EZH2 knockdown was then assessed by real-time quantitative PCR and Western Blot analysis .\n\nCell proliferation was assessed using the soluble tetrazolium salt ( MTS ) cell proliferation assay , and cell cycle was measured by FACS test .\n\nThe methylation status of p16(INK4a )was determined by bisulphate treatment of the DNA , followed by MSP .\n\nEZH2 was over-expressed in NPC cells , and the expression in undifferentiated-derived NPC cells ( CNE-1 , C666-1 ) was more significant than differentiated-derived NPC cells ( HK-1 , CNE-2 ) .\n\nEZH2 was successfully depleted after retroviral infection in C666-1 cells , and the EZH2 depletion could inhibit the proliferation and arrested G1/S phase of NPC cells .\n\nIn addition , both mRNA and protein levels of p16(INK4a) increased significantly in presence of EZH2 depletion .\n\nThe further Methylation-Specific Polymerase Chain Reaction ( MSP ) assay suggested that over-expressed EZH2 may contribute to the reduction of p16(INK4a) expression by hyper methylating its promoter .\n\nEZH2 is overexpressed in NPC and reduces expression of p16(INK4a) by influencing methylation , opening therapeutic options .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "Immunosuppressive oligodeoxynucleotides ( SupODNs ) containing repetitive TTAGGG motifs reduce inflammation and , thus , may have an impact on inflammation\u2011related tumor growth .\n\nIn this study , we found a significant antiproliferative effect of Sup ODNs on the A549 non\u2011small cell lung cancer ( NSCLC ) cell line compared to those treated with control ODNs ( p<0.05 ) .\n\nSup-ODN-mediated G1 phase cell cycle arrest was achieved via inhibition of Akt and extracellular signal-regulated kinase 1/2 phosphorylation and the p15INK4b and p27KIP1/retinoblastoma protein pathway .\n\nIn addition , Sup ODNs induced apoptosis and enhanced apoptosis when combined with vinorelbine .\n\nIn a setting similar to clinical use of multidrug chemotherapy for advanced NSCLC , these effects were investigated by using Sup ODNs in combination with conventional anticancer drugs .\n\nSup ODNs had a significant synergistic effect with 5-fluorouracil , vinorelbine , gemcitabine , paclitaxel and irinotecan , with a mean combination index of 0.43-0.78 ( <1.0 indicates synergism ) in the A549 NSCLC cell line .\n\nIn conclusion , our results showed that Sup ODNs have an anticancer effect and increase the sensitivity of NSCLC cells to conventional anticancer drugs by modifying Akt and the extracellular signal-regulated kinase 1/2 pathway .\n\nThus , Sup ODNs may serve as a novel therapeutic strategy for NSCLC patients .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "In mammalian cells more than 90% of double-strand breaks are repaired by NHEJ .\n\nImpairment of this pathway is associated with cell cycle arrest , cell death , genomic instability and cancer .\n\nHuman diseases such as Nijmegen breakage syndrome , due to mutations in the NBS1 gene , produce defects in resection of double-strand breaks .\n\nNBS1 hypomorphic mutant mice are viable , and cells from these mice are defective in S phase and G2/M checkpoints .\n\nNBS1 polymorphisms have been associated with increased risk of breast cancer .\n\nWe previously demonstrated that estradiol protected estrogen receptor ( ER)-positive ( + ) breast cancer cell lines against double-strand breaks and cell death .\n\nWe now demonstrate that protection from double-strand break damage in ER+ cells is mediated via regulation by c-myc , p53 , CBP and SRC1 coactivators in intron 1 of the NBS1 gene .\n\nWe concluded that NBS1 is responsible for estradiol-mediated protection from double-strand breaks in ER+ breast cancer cells .", "output": "Genomic instability and mutation" }, { "input": "Diarylquinoline compounds are newly synthesized derivatives of the new anti-tuberculosis drug , TMC207 .\n\nIn this study , nine diarylquinoline compounds were screened for cytotoxic activity against human tumor cells , and their mechanisms of action were investigated .\n\nAmong the nine compounds , STM-57 [ N-((6-bromo-2-methoxyquinolin-3-yl)(phenyl)methl)-N-(3,4-dichlorophenyl)-3-(4 -methylpiperazin-1-yl)propanamide ] showed potent cytotoxic activity .\n\nSTM-57 induced caspase-independent cell death in the human nasopharyngeal carcinoma cell line , CNE-2 .\n\nFurther investigation showed that STM-57 induced autophagy , as determined with the increased expression of green fluorescent protein-light chain3 ( GFP-LC3 ) and increased LC3-II levels .\n\nSTM-57 inhibited the phosphorylation of Akt and the mammalian target of rapamycin ( mTOR ) in CNE-2 cells .\n\nThe intracellular calcium concentration and reactive oxygen species levels were increased in CNE-2 cells following treatment with STM-57 , whereas the mitochondrial transmembrane potential ( \u0394\u03a8m ) and ATP concentrations were decreased .", "output": "Tumor promoting inflammation, Resisting cell death" }, { "input": "Here , we provide the necessary proof of concept , that it is possible to metabolically create a non-permissive or \" hostile \" stromal microenvironment , which actively prevents tumor engraftment in vivo .\n\nWe developed a novel genetically engineered fibroblast cell line that completely prevents tumor formation in mice , with a 100% protection rate .\n\nNo host side effects were apparent.This could represent a viable cellular strategy for preventing and treating a variety of human cancers .\n\nMore specifically , we examined the autocrine and paracrine effects of the cellular delivery of TNF-alpha on breast cancer tumor growth and cancer metabolism .\n\nFor this purpose , we recombinantly overexpressed TNF-alpha in human breast cancer cells ( MDA-MB-231 ) or human immortalized fibroblasts ( hTERT-BJ1 ) .\n\nOur results directly show that TNF-alpha functions as a potent tumor suppressor .\n\nRemarkably , TNF-alpha-expressing breast cancer cells were viable , without any significant increases in their basal apoptotic rate .\n\nHowever , after 4 weeks post-implantation , TNF-alpha-expressing breast cancer cells failed to form any tumors in xenografted mice ( 0 tumors/10 injections ) , ultimately conferring 100% protection against tumorigenesis .\n\nSimilarly , TNF-alpha-overexpressing fibroblasts were also viable , without any increases in apoptosis .\n\nSignificantly , complete tumor suppression was obtained by co-injecting TNF-alpha expressing stromal fibroblasts with human breast cancer cells , indicating that paracrine cell-mediated delivery of TNF-alpha can also prevent tumor engraftment and growth ( 0 tumors/10 injections ) .\n\nMechanistically , TNF-alpha induced autophagy and mitochondrial dysfunction in both epithelial cancer cells and stromal fibroblasts , preventing energy transfer from the tumor microenvironment , likely \" starving \" the cancer cells to death .\n\nIn addition , via qRT-PCR analysis of MDA-MB-231 cells , we observed that TNF-alpha mediated the up-regulation of gene transcripts associated with inflammation and senescence [ IL-1-beta , IL-6 , IL-8 , MCP-1 , COX-2 , p21(WAF1/CIP1) ] and downregulated known tumor-promoting genes ( collagen VI and MMP2 ) .\n\nRecombinant overexpression of TNF-alpha receptor(s) in MDA-MB-231 cells also significantly reduced tumor growth , but was not as effective as the TNF-alpha ligand itself in preventing tumor growth .\n\nThus , we propose that stromal cell-mediated delivery of TNF-alpha to human tumors [ using transfected fibroblasts or mesenchymal stem cells ( hMSCs) ] may be a novel and effective strategy for the prevention and treatment of human cancers .", "output": "Tumor promoting inflammation, Enabling replicative immortality, Resisting cell death" }, { "input": "The aberrant expression of human epidermal growth factor receptor-2 ( HER-2 ) has been detected in ovarian cancer .\n\nHowever , the role of HER-2 in the development of ovarian cancer has not been sufficiently elucidated .\n\nThe objective of this study was to determine the role of HER-2 in the apoptosis and metastasis of SKOV-3 ovarian cancer cells .\n\nSKOV-3 cells were transfected with three double\u2011stranded small interfering RNA ( siRNA ) molecules that target HER-2 .\n\nVarious sequences were synthesized by T7 transcription invitro to select the most effective HER-2\u2011silencing siRNA .\n\nSKOV-3 cells were examined for growth inhibition using the MTT proliferation assay and apoptosis was assessed using flow cytometry and TUNEL assay .\n\nThe Matrigel basement memebrane matrix was used to assess invasion and chemotactic mobility , as a model of tumor cell metastasis .\n\nWestern blot analysis was used to detect the expression of matrix metallopeptidase-9 ( MMP-9 ) , E-cadherin , N-cadherin and vimentin. siRNA interference in HER-2 resulted in decreased cell proliferation and invasion and increased apoptosis .\n\nWestern blot analysis demonstrated a marked increase in E-cadherin and MMP-9 and a reduction in N-cadherin and vimentin protein levels in the SKOV-3 cells .\n\nThe suppression of HER-2 expression resulted in apoptosis and the inhibition of metastasis of SKOV-3 cells .\n\nTherefore , the overexpression of the HER-2 gene can enhance the metastatic potential of SKOV-3 cells by increasing the protein levels of MMP-9 .\n\nEpithelial-mesenchymal transition may be involved in the HER-2 siRNA-induced invasion and migration of SKOV-3 cells .\n\nTaken together , these results suggest that HER-2 functions as an oncogene and may thus be an attractive therapeutic target in SKOV-3 ovarian cancer cells .", "output": "Sustaining proliferative signaling, Resisting cell death, Activating invasion and metastasis" }, { "input": "Cell invasion is required for neoplastic metastasis .\n\nMatrix metalloproteinase-9 ( MMP-9 ) , which degrades the extracellular matrix , is a major component in the process of cancer cell invasion .\n\nSulfuretin is one of the major flavonoids isolated from Rhus verniciflua .\n\nSulfuretin has been used to reduce oxidative stress , platelet aggregation , the inflammatory response and mutagenesis .\n\nHowever , the effect of sulfuretin on breast cancer metastasis is unknown .\n\nIn this study , we investigated the inhibitory effect of sulfuretin on 12-O-tetradecanoylphorbol-13-acetate ( TPA)-induced MMP-9 expression and cell invasion in MCF-7 cells .\n\nSulfuretin inhibited TPA-induced transcriptional activation of nuclear factor-\\u03baB ( NF-\\u03baB ) .\n\nWe demonstrated that sulfuretin mediated the inhibition of TPA-induced MMP-9 expression and that cell invasion in MCF-7 cells involved suppression of the NF-\\u03baB pathway .\n\nTherefore , inhibiting MMP-9 expression by sulfuretin may have therapeutic potential for controlling breast cancer invasiveness .", "output": "Activating invasion and metastasis" }, { "input": "Prostate cancer is responsible for major deaths globally after lung cancer .\n\nHowever , etiology of prostate cancer is still unknown .\n\nIndividual risk and incidence of prostate cancer may result from the interaction of genetic susceptibility with exposure to environmental factors such as infectious agents , tobacco , occupational exposure , dietary carcinogens , and/or hormonal imbalances leading to injury of the prostate and to the development of chronic inflammation .\n\nAbout 30% of all human cancers are caused by tobacco smoking and inhaled pollutants .\n\nInflammation is now regarded as an important hallmark of cancer .\n\nThe present study has been aimed to explore the pro-inflammatory levels in prostate carcinoma patients by examining the serum levels of novel cytokine interleukin-18 ( IL-18 ) expression in tobacco exposed population .\n\nA total of 578 ( n = 284 biopsy proven prostate cancer patients , n = 294 controls with and without tobacco exposed population ) were recruited .\n\nSerum IL-18 ( Interleukin-18 ) level was done by ELISA .\n\nThe IL-18 levels between cancer patients and controls within same mode tobacco exposure as tobacco smoking ( overall ) showed significant difference ( P < 0.0001 ) and further we compared within stratified group , it significantly differ ( P < 0.0001 ) in bidi and cigarette smoking than control non users .\n\nFurthermore , IL-18 levels in tobacco chewers ( overall ) with gutkha and khaini chewers showed significant difference ( P < 0.01 ) than controls non users .\n\nMoreover , the IL-18 levels between cancer patients and controls with in of combined mode chewers smokers and alcohol ( CSA ) , smokers with alcohol showed significant difference ( P < 0.01 ) than controls .\n\nThe IL-18 levels also differed significantly ( P < 0.05 ) with smokers and chewers in higher stages of III and IV , and showed non significant with in lower stages .\n\nTobacco exposure enhance the inflammation in prostate carcinoma patients in stratified group as it have been represented as a risk factors in various cancers , but this study provide further its role that seems to influence inflammation especially in prostate carcinoma .", "output": "Tumor promoting inflammation" }, { "input": "OBJECTIVE In addition to its effects on cholesterol levels , apoE3 has lipid-independent effects that contribute to cardiovascular protection ; one of these effects is the ability to inhibit cell cycling in VSMCs .\n\nThe goal of this study was to identify and characterize cell cycle-regulatory mechanisms responsible for the anti-mitogenic effect of apoE .\n\nMETHODS AND RESULTS Primary VSMCs were stimulated with serum in the absence or presence of apoE3. apoE3 upregulated expression of the cdk inhibitor , p27(kip1) , in primary VSMCs , and this effect required Cox2 and activation of PGI(2)-IP signaling .\n\nThe microRNA family , miR221/222 has recently been identified as a post-translational regulator of p27 , and apoE3 inhibited miR221/222 expression in a Cox2- and PGI(2)/IP-dependent manner .\n\nMoreover , reconstituted miR222 expression was sufficient to override the effects of apoE on p27 expression and S phase entry .\n\nThe ability to repress expression of miR221/222 is shared by apoE3-containing HDL but is absent from apoA-1 , LDL and apoE-depleted HDL .\n\nAll three apoE isoforms regulate miR221/222 , and the effect is independent of the C-terminal lipid-binding domain. miR221/222 levels are increased in the aortae of apoE3-null mice and reduced when apoE3 expression is reconstituted by adeno-associated virus infection .\n\nThus , regulation of miR221/222 by apoE3 occurs invivo as well as invitro .\n\nCONCLUSIONS ApoE inhibits VSMC proliferation by regulating p27 through miR221/222 .\n\nControl of cell cycle-regulatory microRNAs adds a new dimension to the spectrum of cardiovascular protective effects afforded by apoE and apoE-HDL .", "output": "Evading growth suppressors" }, { "input": "Protein kinase B ( AKT)/mammalian target of rapamycin ( mTOR ) signaling pathway plays a crucial role in the tumorigenesis and progression of multiple tumors , and has been shown to be important therapeutic targets for cancer .\n\nThe present study aimed to explore the role and molecular mechanisms of AKT/mTOR pathway in human hemangioma ( HA ) .\n\nTwenty-five cases of human HA tissues were collected .\n\nThe expression of AKT , mTOR and proliferating cell nuclear antigen ( PCNA ) proteins was evaluated using semi-quantitative immunohistochemistry in biopsy samples in different phases of HA .\n\nAKT/mTOR pathway was blocked by recombinant small hairpin RNA adenovirus vector rAd5-AKT+mTOR ( rAd5-Am ) , used for infecting proliferating phase HA-derived endothelial cells ( HDEC ) .\n\nThe expression of AKT , mTOR and PCNA was detected by Real-time PCR and Western blot assays .\n\nCell proliferative activities were determined by MTT assay , and cell cycle distribution and apoptosis were analyzed by flow cytometry .\n\nAs a consequence , the expression of AKT , mTOR and PCNA was significantly increased in proliferative phase HA , while that was decreased in involutive phase .\n\nCombined blockade of AKT/mTOR pathway by rAd5-Am diminished cell proliferative activities , and induced cell apoptosis and cycle arrest with the decreased expression of AKT , mTOR and PCNA in proliferative phase HDEC .\n\nIn conclusion , the activity of AKT/mTOR pathway was increased in proliferative phase HA , while it was decreased in involutive phase .\n\nCombined blockade of AKT/mTOR pathway might suppress cell proliferation via down-regulation of PCNA expression , and induce apoptosis and cycle arrest in proliferative phase HDEC , suggesting that AKT/mTOR pathway might represent the important therapeutic targets for human HA .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "Ovarian cancer , one of inflammation-associated cancers , is the fifth leading cause of cancer deaths among women .\n\nInflammation in the tumor microenvironment is associated with peritoneal tumor dissemination and massive ascites , which contribute to high mortality in ovarian cancer .\n\nTumor suppressor p53 is frequently deleted or mutated in aggressive and high-grade ovarian cancer , probably aggravating cancer progression and increasing mortality .\n\nWe therefore investigated the influence of p53 on proinflammatory chemokines in ovarian cancer cells .\n\nA PCR array of the chemokine network revealed that ovarian cancer cells with low or mutated p53 expression expressed high levels of proinflammatory chemokines such as CXCL1 , 2 , 3 and 8 .\n\nTransient transfection of p53 into p53-null ovarian cancer cells downregulated proinflammatory chemokines induced by tumor necrosis factor-\u03b1 ( TNF ) , a proinflammatory cytokine abundantly expressed in ovarian cancer .\n\nFurthermore , p53 restoration or stabilization blocked TNF-induced NF-\u03baB promoter activity and reduced TNF-activated I\u03baB .\n\nRestoration of p53 increased ubiquitination of I\u03baB , resulting from concurrently reduced proteasome activity followed by stability of I\u03baB .\n\nA ubiquitination PCR array on restoration of p53 did not reveal any significant change in expression except for Mdm2 , indicating that the balance between p53 and Mdm2 is more important in regulating NF-\u03baB signaling rather than the direct effect of p53 on ubiquitin-related genes or I\u03baB kinases .\n\nIn addition , nutlin-3 , a specific inducer of p53 stabilization , inhibited proinflammatory chemokines by reducing TNF-activated I\u03baB through p53 stabilization .\n\nTaken together , these results suggest that p53 inhibits proinflammatory chemokines in ovarian cancer cells by reducing proteasomal degradation of I\u03baB .\n\nThus , frequent loss or mutation of p53 may promote tumor progression by enhancing inflammation in the tumor microenvironment .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "Approximately half of all HER2/neu-overexpressing breast cancer patients do not respond to trastuzumab-containing therapy .\n\nTherefore , there remains an urgent and unmet clinical need for the development of predictive biomarkers for trastuzumab response .\n\nRecently , several lines of evidence have demonstrated that the inflammatory tumor microenvironment is a major contributor to therapy resistance in breast cancer .\n\nIn order to explore the predictive value of inflammation in breast cancer patients , we measured the inflammatory biomarkers serum ferritin and C-reactive protein ( CRP ) in 66 patients immediately before undergoing trastuzumab-containing therapy and evaluated their progression-free and overall survival .\n\nThe elevation in pre-treatment serum ferritin ( >250 ng/ml ) or CRP ( >7.25 mg/l ) was a significant predictor of reduced progression-free survival and shorter overall survival .\n\nWhen patients were stratified based on their serum ferritin and CRP levels , patients with elevation in both inflammatory biomarkers had a markedly poorer response to trastuzumab-containing therapy .\n\nTherefore , the elevation in inflammatory serum biomarkers may reflect a pathological state that decreases the clinical efficacy of this therapy .\n\nAnti-inflammatory drugs and life-style changes to decrease inflammation in cancer patients should be explored as possible strategies to sensitize patients to anti-cancer therapeutics .", "output": "Tumor promoting inflammation" }, { "input": "Nuclear factor kappa-B ( NF-\u03baB ) activates multiple genes with overlapping roles in cell proliferation , inflammation and cancer .\n\nUsing an unbiased approach we identified human CDK6 as a novel kinase phosphorylating NF-\u03baB p65 at serine 536 .\n\nPurified and reconstituted CDK6/cyclin complexes phosphorylated p65 in vitro and in transfected cells .\n\nThe physiological role of CDK6 for basal as well as cytokine-induced p65 phosphorylation or NF-\u03baB activation was revealed upon RNAi-mediated suppression of CDK6 .\n\nInhibition of CDK6 catalytic activity by PD332991 suppressed activation of NF-\u03baB and TNF-induced gene expression .\n\nIn complex with a constitutively active viral cyclin CDK6 stimulated NF-\u03baB p65-mediated transcription in a target gene specific manner and this effect was partially dependent on its ability to phosphorylate p65 at serine 536 .\n\nTumor formation in thymi and spleens of v-cyclin transgenic mice correlated with increased levels of p65 Ser536 phosphorylation , increased expression of CDK6 and upregulaton of the NF-\u03baB target cyclin D3 .\n\nThese results suggest that aberrant CDK6 expression or activation that is frequently observed in human tumors can contribute through NF-\u03baB to chronic inflammation and neoplasia .", "output": "Tumor promoting inflammation" }, { "input": "Activation of p53 effectively inhibits tumor angiogenesis that is necessary for tumor growth and metastasis .\n\nReactivation of the p53 by small molecules has emerged as a promising new strategy for cancer therapy .\n\nSeveral classes of small-molecules that activate the p53 pathway have been discovered using various approaches .\n\nHere , we identified harmine ( \u03b2-carboline alkaloid ) as a novel activator of p53 signaling involved in inhibition of angiogenesis and tumor growth .\n\nHarmine induced p53 phosphorylation and disrupted the p53-MDM2 interaction .\n\nHarmine also prevented p53 degradation in the presence of cycloheximide and activated nuclear accumulation of p53 followed by increasing its transcriptional activity in endothelial cells .\n\nMoreover , harmine not only induced endothelial cell cycle arrest and apoptosis , but also suppressed endothelial cell migration and tube formation as well as induction of neovascularity in a mouse corneal micropocket assay .\n\nFinally , harmine inhibited tumor growth by reducing tumor angiogenesis , as demonstrated by a xenograft tumor model .\n\nOur results suggested a novel mechanism and bioactivity of harmine , which inhibited tumor growth by activating the p53 signaling pathway and blocking angiogenesis in endothelial cells .", "output": "Inducing angiogenesis, Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "Angiogenesis , the formation of new blood vessels from pre-existing vascular beds , is essential for tumor growth , invasion , and metastasis .\n\nLuteolin is a common dietary flavonoid found in fruits and vegetables .\n\nWe studied the antiangiogenic activity of luteolin using in vitro , ex vivo , and in vivo models .\n\nIn vitro studies using rat aortic ring assay showed that luteolin at non-toxic concentrations significantly inhibited microvessel sprouting and proliferation , migration , invasion and tube formation of endothelial cells , which are key events in the process of angiogenesis .\n\nLuteolin also inhibited ex vivo angiogenesis as revealed by chicken egg chorioallantoic membrane assay ( CAM ) and matrigel plug assay .\n\nGelatin zymographic analysis demonstrated the inhibitory effect of luteolin on the activation of matrix metalloproteinases MMP-2 and MMP-9 .\n\nWestern blot analysis showed that luteolin suppressed VEGF induced phosphorylation of VEGF receptor 2 and their downstream protein kinases AKT , ERK , mTOR , P70S6K , MMP-2 , and MMP-9 in HUVECs .\n\nProinflammatory cytokines such as IL-1\u03b2 , IL-6 , IL-8 , and TNF-\u03b1 level were significantly reduced by the treatment of luteolin in PC-3 cells .\n\nLuteolin ( 10 mg/kg/d ) significantly reduced the volume and the weight of solid tumors in prostate xenograft mouse model , indicating that luteolin inhibited tumorigenesis by targeting angiogenesis .\n\nCD31 and CD34 immunohistochemical staining further revealed that the microvessel density could be remarkably suppressed by luteolin .\n\nMoreover , luteolin reduced cell viability and induced apoptosis in prostate cancer cells , which were correlated with the downregulation of AKT , ERK , mTOR , P70S6K , MMP-2 , and MMP-9 expressions .\n\nTaken together , our findings demonstrate that luteolin inhibits human prostate tumor growth by suppressing vascular endothelial growth factor receptor 2-mediated angiogenesis .", "output": "Inducing angiogenesis, Resisting cell death" }, { "input": "Tumors are infiltrated by macrophages , T and B-lymphocytes , which may favor tumor development by promoting angiogenesis , growth and invasion .\n\nThe aim of this study was to investigate the clinical relevance of the relative amount of macrophages ( CD68\u207a ) , T-cells ( CD3\u207a and B-cells ( CD20\u207a ) at the invasive front of breast carcinomas , and the expression of matrix metalloproteases ( MMPs ) and their inhibitors ( TIMPs ) either at the invasive front or at the tumor center .\n\nWe performed an immunohistochemical study counting CD3 , CD20 and CD68 positive cells at the invasive front , in 102 breast carcinomas .\n\nAlso , tissue sections were stained with MMP-2 , -9 , -11 , -14 and TIMP-2 antibodies , and immunoreactivity location , percentage of reactive area and intensity were determined at the invasive front and at the tumor center .\n\nThe results showed that an increased CD68 count and CD68/(CD3+CD20) ratio were directly associated with both MMP-11 and TIMP-2 expression by mononuclear inflammatory cells at the tumor center ( p\u200a=\u200a0.041 and p\u200a=\u200a0.025 for CD68 count and p\u200a=\u200a0.001 and p\u200a=\u200a0.045 for ratio , respectively for MMP-11 and TIMP-2 ) .\n\nIn addition , a high CD68/(CD3+CD20) ratio ( >0.05 ) was directly associated with a higher probability of shortened relapse-free survival .\n\nMultivariate analysis revealed that CD68/(CD3+CD20) ratio was an independent factor associated with distant relapse-free survival ( RR : 2.54 , CI : ( 1.23-5.24 ) , p<0.01 ) .\n\nTherefore , CD68/(CD3+CD20) ratio at the invasive front could be used as an important prognostic marker .", "output": "Tumor promoting inflammation, Avoiding immune destruction" }, { "input": "Lung cancer is the leading cause of cancer-related deaths in the world .\n\nAchaete-scute complex homolog-1 ( Ascl1 ) is a member of the basic helix-loop-helix ( bHLH ) transcription factor family that has multiple functions in the normal and neoplastic lung such as the regulation of neuroendocrine differentiation , prevention of apoptosis and promotion of tumor-initiating cells .\n\nWe now show that Ascl1 directly regulates matrix metalloproteinase-7 ( MMP-7 ) and O(6)-methylguanine-DNA methyltransferase ( MGMT ) .\n\nLoss- and gain-of-function experiments in human bronchial epithelial and lung carcinoma cell lines revealed that Ascl1 , MMP-7 and MGMT are able to protect cells from the tobacco-specific nitrosamine NNK-induced DNA damage and the alkylating agent cisplatin-induced apoptosis .\n\nWe also examined the role of Ascl1 in NNK-induced lung tumorigenesis in vivo .\n\nUsing transgenic mice which constitutively expressed human Ascl1 in airway lining cells , we found that there was a delay in lung tumorigenesis .\n\nWe conclude that Ascl1 potentially enhances DNA repair through activation of MMP-7 and MGMT which may impact lung carcinogenesis and chemoresistance .\n\nThe study has uncovered a novel and unexpected function of Ascl1 which will contribute to better understanding of lung carcinogenesis and the broad implications of transcription factors in tobacco-related carcinogenesis .", "output": "Genomic instability and mutation, Resisting cell death" }, { "input": "We had demonstrated that plasminogen kringle 5 ( K5 ) , a potent angiogenic inhibitor , inhibited retinal neovascularization and hepatocellular carcinoma growth by anti-angiogenesis .\n\nThe current study investigated the effects and the underlying mechanisms of K5 on both tumor growth and spontaneous pulmonary metastasis in Lewis lung carcinoma ( LLC ) implanted mouse model .\n\nSimilarly , K5 could decrease expression of VEGF in LLC cells and grafted tissues and suppress tumor angiogenesis and growth .\n\nK5 had no direct effect on proliferation and apoptosis of LLC .\n\nHowever , K5 could significantly inhibit SDF-1\u03b1-induced chemotaxis movement of LLC cells and resulted in a great reduction of surface metastatic nodules and micrometastases in the lungs of LLC tumor-bearing mice .\n\nK5 also decreased expression of chemokine ( C-X-C motif ) receptor 4 ( CXCR4 ) in LLC cells and grafted tissues .\n\nFurthermore , K5 down-regulated SDF-1\u03b1 expression in metastatic lung tissues of LLC-bearing mice .\n\nTherefore , K5 may suppress tumor pulmonary metastasis through inhibiting SDF-1\u03b1-CXCR4 chemotaxis movement and down-regulation of VEGF .\n\nMoreover , the role of hypoxia inducible factor-1\u03b1 ( HIF-1\u03b1 ) , a crucial transcriptional factor for both VEGF and CXCR4 expression , was evaluated .\n\nThe siRNA of HIF-1\u03b1 attenuated expression of VEGF and CXCR4 and inhibited LLC migration .\n\nK5 decreased HIF-1\u03b1 protein level and impaired nuclear HIF-1\u03b1 accumulation .\n\nThese results showed for the first time that K5 inhibits LLC growth and metastasis via the dual effects of anti-angiogenesis and suppression of tumor cell motility by targeting the pivotal molecule , HIF-1\u03b1 .", "output": "Activating invasion and metastasis, Inducing angiogenesis, Sustaining proliferative signaling, Resisting cell death" }, { "input": "Novel strategies are necessary to improve chemotherapy response in advanced and recurrent endometrial cancer .\n\nHere , we demonstrate that terpenoids present in the Steam Distilled Extract of Ginger ( SDGE ) are potent inhibitors of proliferation of endometrial cancer cells .\n\nSDGE , isolated from six different batches of ginger rhizomes , consistently inhibited proliferation of the endometrial cancer cell lines Ishikawa and ECC-1 at IC(50) of 1.25 \ufffdg/ml .\n\nSDGE also enhanced the anti-proliferative effect of radiation and cisplatin .\n\nDecreased proliferation of Ishikawa and ECC-1 cells was a direct result of SDGE-induced apoptosis as demonstrated by FITC-Annexin V staining and expression of cleaved caspase 3 .\n\nGC/MS analysis identified a total of 22 different terpenoid compounds in SDGE , with the isomers neral and geranial constituting 30-40% .\n\nCitral , a mixture of neral and geranial inhibited the proliferation of Ishikawa and ECC-1 cells at an IC(50) 10 \ufffdM ( 2.3 \ufffdg/ml ) .\n\nPhenolic compounds such as gingerol and shogaol were not detected in SDGE and 6-gingerol was a weaker inhibitor of the proliferation of the endometrial cancer cells .\n\nSDGE was more effective in inducing cancer cell death than citral , suggesting that other terpenes present in SDGE were also contributing to endometrial cancer cell death .\n\nSDGE treatment resulted in a rapid and strong increase in intracellular calcium and a 20-40% decrease in the mitochondrial membrane potential .\n\nSer-15 of p53 was phosphorylated after 15 min treatment of the cancer cells with SDGE .\n\nThis increase in p53 was associated with 90% decrease in Bcl2 whereas no effect was observed on Bax .\n\nInhibitor of p53 , pifithrin-\u03b1 , attenuated the anti-cancer effects of SDGE and apoptosis was also not observed in the p53(neg) SKOV-3 cells .\n\nOur studies demonstrate that terpenoids from SDGE mediate apoptosis by activating p53 and should be therefore be investigated as agents for the treatment of endometrial cancer .", "output": "Resisting cell death" }, { "input": "Hormone antagonist therapy for estrogen receptor positive ( ER+ ) breast cancer patients post radical surgery and radiation therapy has a poor prognosis and also causes bone loss. 1\u03b1,25-dihydroxyvitamin D(3) [ 1\u03b1,25(OH)(2)D(3) ] is a potent antitumor agent in pre-clinical studies , but caused hypercalcemia when its effective antitumor doses were used .\n\nTherefore , we investigated the effects of a less-calcemic 1\u03b1,25(OH)(2)D(3) analog , 19-nor-2\u03b1-(3-hydroxypropyl)-1\u03b1,25-dihydroxyvitamin D(3 )(MART-10 ) , on ER+MCF-7 cells .\n\nWe demonstrate that MART-10 is 500- to 1000-fold more potent than 1\u03b1,25(OH)(2)D(3) in inhibiting cell growth in a dose- and time-dependent manner .\n\nMART-10 is also much more potent in arresting MCF-7cell cycle progression at G(0)/G(1) phase as compared to 1\u03b1,25(OH)(2)D(3) , possibly mediated by a greater induction of p21 and p27 expression .\n\nMoreover , MART-10 is more active than 1\u03b1,25(OH)(2)D(3) in causing cell apoptosis , likely through a higher BAX/Bcl expression ratio and the subsequent cytochrome C release from mitochondria to cytosol .\n\nBased on our in vitro findings , MART-10 could be a promising vitamin D analog for the potential treatment of breast cancer , for example , ER+ patients , to decrease the tumor relapse rate and the side effect on bone caused by antihormone regimens .\n\nThus , further in vivo animal study is warranted .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "Retinoic acid ( RA ) has been believed to be an anticancer drug for a long history .\n\nHowever , the molecular mechanisms of RA actions on cancer cells remain diverse .\n\nIn this study , the dose-dependent inhibition of RA on DU145 cell proliferation was identified .\n\nInterestingly , RA treatment triggered p35 cleavage ( p25 formation ) and Cdk5 overactivation , and all could be blocked by Calpain inhibitor , Calpeptin ( CP ) .\n\nSubsequently , RA-triggered DU145 apoptosis detected by sub-G1 phase accumulation and Annexin V staining could also be blocked by CP treatment .\n\nFurthermore , RA-triggered caspase 3 activation and following Cdk5 over-activation were destroyed by treatments of both CP and Cdk5 knockdown .\n\nIn conclusion , we report a new mechanism in which RA could cause apoptosis of androgen-independent prostate cancer cells through p35 cleavage and Cdk5 over-activation .\n\nThis finding may contribute to constructing a clearer image of RA function and bring RA as a valuable chemoprevention agent for prostate cancer patients .", "output": "Resisting cell death" }, { "input": "Background .\n\nMultiple myeloma ( MM ) , an almost incurable disease , is the second most common blood cancer .\n\nInitial chemotherapeutic treatment could be successful ; however , resistance development urges the use of higher toxic doses accompanied by hematopoietic stem cell transplantation .\n\nThe establishment of more effective treatments that can overcome or circumvent chemoresistance has become a priority .\n\nWe recently demonstrated that venom extracted from Walterinnesia aegyptia ( WEV ) either alone or in combination with silica nanoparticles ( WEV+NPs ) mediated the growth arrest and apoptosis of prostate cancer cells .\n\nIn the present study , we evaluated the impact of WEV alone and WEV+NP on proliferation and apoptosis of MM cells .\n\nMethods .\n\nThe impacts of WEV alone and WEV+NP were monitored in MM cells from 70 diagnosed patients .\n\nThe influences of WEV and WEV+NP were assessed with flow cytometry analysis .\n\nResults .\n\nWEV alone and WEV+NP decreased the viability of MM cells .\n\nUsing a CFSE proliferation assay , we found that WEV+NP strongly inhibited MM cell proliferation .\n\nFurthermore , analysis of the cell cycle using the propidium iodide ( PI ) staining method indicated that WEV+NP strongly altered the cell cycle of MM cells and enhanced the induction of apoptosis .\n\nConclusions .\n\nOur data reveal the biological effects of WEV and WEV+NP on MM cells that enable these compounds to function as effective treatments for MM .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "Background and Objective .\n\nThe cell cycle is regulated by proteins at different checkpoints , and dysregulation of this cycle plays a role in carcinogenesis .\n\nMatrix metalloproteinases ( MMPs ) are enzymes that degrade collagen and promote tumour infiltration .\n\nThe aim of this study was to evaluate the expression of various cell cycle regulators and MMPs and to correlate such expression with progression and recurrence in patients with stage T1 urothelial carcinoma of the bladder ( UCB ) .\n\nPatients and Methods .\n\nThis population-based cohort study comprised 201 well-characterized patients with primary stage T1 urothelial carcinoma of the bladder .\n\nImmunohistochemistry was performed on formalin-fixed material to quantify expression of cell cycle regulators and two MMPs .\n\nResults .\n\nNormal expression of p53 and abnormal expression of MMP9 were associated with greater risk of tumour recurrence .\n\nAlso , normal p16 expression was related to a lower risk of tumour progression .\n\nMMP2 , p21 , cyclin D1 , and pRb showed no significant results that could estimate progression or recurrence .\n\nConclusions .\n\nNormal p16 expression is associated with a lower risk of tumour progression , but immunohistochemistry on cell cycle regulators and MMPs has little value in predicting the prognosis in stage T1 UCB .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "The expression of the angiogenic phenotype is regulated by a balance of pro-angiogenic and anti-angiogenic factors released into the tumor microenvironment .\n\nNuclear protein 7 ( NOL7 ) , a novel tumor suppressor , acts as a master regulator of angiogenesis by downregulating pro-angiogenic factors and upregulating anti-angiogenic factors .\n\nUsing cervical cancer as a model of investigation , we have previously shown that loss of NOL7 mRNA and protein expression is observed as early as the premalignant phase .\n\nAnalysis of the gene failed to identify tumor-specific promoter methylation or coding region mutations , suggesting that NOL7 loss may be mediated by aberrant expression of its upstream regulators .\n\nIn this study , we show that the RB tumor suppressor gene ( RB ) positively regulates NOL7 at the transcriptional level by recruiting transcription factors and transcription machinery proteins to its promoter region .\n\nConversely , the loss of RB represses NOL7 transcription by inhibiting assembly of these proteins .\n\nThis loss of NOL7 expression is also observed in RB-deficient human malignancies .\n\nTogether , this work further characterizes the transcriptional activator function of RB and defines a potential role for RB in regulating angiogenesis through activation of NOL7 .\n\nCurrent anti-angiogenic therapies lack long-term efficacy , as they are unable to target the diverse angiogenic signals generated by tumors .\n\nOur data provide evidence to support the hypothesis that reactivation of pRB can potentially modulate the expression of the angiogenic phenotype through regulation of NOL7 .\n\nTherefore , this knowledge may be employed to design more comprehensive and effective therapies .", "output": "Inducing angiogenesis" }, { "input": "Luteolin is a plant flavonoid which exhibits anti-oxidative , anti-inflammatory and anti-tumor effects .\n\nHowever , the antiproliferative potential of luteolin is not fully understood .\n\nIn this study , we investigated the effect of luteolin on cell cycling and apoptosis in human esophageal squamous carcinoma cell line Eca109 cells .\n\nMTT assays showed that luteolin had obvious cytotoxicity on Eca109 with an IC50 of 70.7\ufffd1.72 \u03bcM at 24 h .\n\nLuteolin arrested cell cycle progression in the G0/G1 phase and prevented entry into S phase in a dose- and time-dependent manner. as assessed by FCM .\n\nLuteolin induced apoptosis of Eca109 cells was demonstrated by AO/EB staining assay and annexin V-FITC/PI staining .\n\nMoreover , luteolin downregulated the expression of cyclin D1 , survivin and c-myc , and it also upregulated the expression of p53 , in line with the fact that luteolin was able to inhibit Eca109 cell proliferation .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "OBJECTIVE The aim of this retrospective study is to analyze the clinical and pathological factors related to the prognosis of Chinese patients with stage Ib to IIb cervical cancer .\n\nMETHODS AND RESULTS 13 clinical pathological factors in 255 patients with stage Ib to IIb cervical cancer undergoing radical hysterectomy and systematic lymphadenectomy were analyzed to screen for factors related to prognosis .\n\nThe cumulative 5-year survival of the 255 patients was 75.7% .\n\nThe result of the univariate analysis suggested that clinical stage , cell differentiation , depth of cervical stromal invasion , parametrial tissue involvement , and lymph node metastasis were prognostic factors for patients with stage Ib to IIb cervical cancer ( P<0.05 ) .\n\nCompared with cases with involvement of iliac nodes , obturator nodes , or inguinal lymph nodes , cases with metastasis to the common iliac lymph nodes had a poorer prognosis ( P<0.05 ) .\n\nCases with involvement of four or more lymph nodes had a poorer prognosis than those with involvement of three or fewer lymph nodes ( P<0.05 ) .\n\nUsing multivariate Cox proportional hazards model regression analysis , non-squamous histological type , poor differentiation , parametrial tissue involvement , and outer 1/3 stromal invasion were found to be independently related to patients poor prognosis ( P<0.05 ) .\n\nCONCLUSION Non-squamous histological type , poor cell differentiation , parametrial tissue involvement , and outer 1/3 stromal invasion are the independent poor prognostic factors for patients with stage Ib to IIb cervical cancer .", "output": "Activating invasion and metastasis" }, { "input": "Onions ( Allium cepa L. ) are widely used in the food industry for its nutritional and aromatic properties .\n\nOur studies showed that ethyl acetate extract of onion ( EEO ) had potent inhibitory effects on animal fatty acid synthase ( FAS ) , and could induce apoptosis in FAS over-expressing human breast cancer MDA-MB-231 cells .\n\nFurthermore , this apoptosis was accompanied by reduction of intracellular FAS activity and could be rescued by 25 mM or 50 mM exogenous palmitic acids , the final product of FAS catalyzed synthesis .\n\nThese results suggest that the apoptosis induced by EEO occurs via inhibition of FAS .\n\nWe also found that EEO could suppress lipid accumulation during the differentiation of 3T3-L1 adipocytes , which was also related to its inhibition of intracellular FAS activity .\n\nSince obesity is closely related to breast cancer and obese patients are at elevated risk of developing various cancers , these findings suggested that onion might be useful for preventing obesity-related malignancy .", "output": "Resisting cell death" }, { "input": "Family with sequence similarity 189 , member B ( FAM189B ) , alias COTE1 , a putative oncogene selected by microarray , for the first time was here found to be significantly up-regulated in hepatocellular carcinoma ( HCC ) specimens and HCC cell lines. mRNA expression of COTE1 in HCC samples and cell lines was detected by reverse transcription-polymerase chain reaction ( RT-PCR ) and real-time PCR , while protein expression of COTE1 in HCC tissues was assessed by immunohistochemistry .\n\nIn addition , invasion of HCC cells was observed after overexpressing or silencing COTE1 .\n\nIn the total of 48 paired HCC specimens , compared with the adjacent non-cancer tissues , the expression of COTE1 was up-regulated in 31 ( p<0.01 ) .\n\nIn HCC cell lines , COTE1 expression was significantly higher than in normal human adult liver ( p<0.01 ) .\n\nOverexpression of COTE1 enhanced HCC-derived LM6 and MHCC-L cellular invasion in vitro .\n\nIn contrast , COTE1 knockdown via RNAi markedly suppressed these phenotypes , as documented in LM3 and MHCC-H HCC cells .\n\nMechanistic analyses indicated that COTE1 could physically associate with WW domain oxidoreductase ( WWOX ) , a tumor suppressor .\n\nCOTE1 may be closely correlated with invasion of hepatocellular carcinoma ( HCC ) cells and thus may serve as an effective target for gene therapy .", "output": "Activating invasion and metastasis" }, { "input": "Breast cancer is the most commonly diagnosed cancer in women in the world and is one of the leading causes of death due to cancer .\n\nHealth benefits have been linked to additive and synergistic combinations of phytochemicals in fruits and vegetables .\n\nNigella sativa has been shown to possess anti-carcinogenic activity , inhibiting growth of several cancer cell lines in vitro .\n\nHowever , the molecular mechanisms of the anti-cancer properties of Nigella sativa phytochemical extracts have not been completely understood .\n\nOur data showed that Nigella sativa extracts significantly inhibited human breast cancer MDA-MB-231 cell proliferation at doses of 2.5-5 \u03bcg/mL ( P<0.05 ) .\n\nApoptotic induction in MDA-MB-231 cells was observed in a dose-dependent manner after exposure to Nigella sativa extracts for 48 h .\n\nReal time PCR and flow cytometry analyses suggested that Nigella sativa extracts possess the ability to suppress the proliferation of human breast cancer cells through induction of apoptosis .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "Momordica charantia has been found to exhibit anticancer activity , in addition to its well-known therapeutic functions .\n\nWe have demonstrated that the leaf extract of Momordica charantia ( MCME ) induces apoptosis in several human cancer cells through caspase- and mitochondria-dependent pathways .\n\nIn this study , a different susceptibility to MCME was found in human lung adenocarcinoma CL1 cells with different metastatic ability , leading to the significant difference of cell viability and invasiveness between MCME-treated CL1-0 and CL1-5 cells .\n\nMCME was found to upregulate the expression of Wnt-2 and affect the migratory and invasive ability of CL1 cells through suppressed MMP-2 and MMP-9 enzymatic activities .\n\nWe proposed that MCME mediates inhibition against migration of CL1 cells by reducing the expression and activation of Src and FAK to decrease the expression of downstream Akt , \u03b2-catenin , and MMPs .", "output": "Activating invasion and metastasis" }, { "input": "Acute lymphoblastic leukemia is still an incurable disease with resistance to therapy developing in the majority of patients .\n\nWe investigated the effect of TPEN , an intracellular zinc chelator , in Jurkat and in ex vivo acute lymphoblastic leukemia ( ALL ) cells resistant to chemotherapy .\n\nChanges of nuclei morphology , reactive oxygen species generation , presence of hypodiploid cells , phosphatidylserine translocation , mitochondrial membrane depolarization , immunohistochemical identification of cell death signalling molecules , and pharmacological inhibition were assayed to detect the apoptotic cell death pathways .\n\nWe found that TPEN induces apoptosis in both types of cells by a molecular oxidative stress pathway involving O(2)(\u2022-) > H(2)O(2) > NF-\u03baB ( JNK/c-Jun ) >p53> loss \u0394\u03a8(m)> caspase-3 , AIF > chromatin condensation/DNA fragmentation .\n\nInterestingly , TPEN induced apoptosis independently of glucose ; leukemic cells are therefore devoid of survival capacity by metabolic resistance to treatment .\n\nMost importantly , TPEN cytotoxic effect can eventually be regulated by the antioxidant N-acetyl-cysteine and zinc ions .\n\nOur data suggest that TPEN can be used as a potential therapeutic prooxidant agent against refractory leukemia .\n\nThese data contribute to understanding the importance of oxidative stress in the treatment of ALL .", "output": "Tumor promoting inflammation, Resisting cell death" }, { "input": "The Punica granatum L. var. granatum ( pomegranate ) has been demonstrated to exert antitumor effects on various types of cancer cells .\n\nThe present study aimed to evaluate the medicinal herbs Punica granatum L. var. spinosa ( apple punice ) that are native to Iran .\n\nThis study was determined to test the possible cytotoxic activity and induction of apoptosis on human prostate cell lines .\n\nThe effect of ethanol extracts of the herbs on the inhibition of cell proliferation was assessed by MTT colorimetric assay .\n\nPC3 cell lines treated with the extracts were analyzed for the induction of apoptosis by cell death detection ( ELISA ) and TUNEL assay .\n\nDye exclusion analysis was performed for viability rate .\n\nOur results demonstrated that the Punica granatum L. var. spinosa extract dose dependently suppressed the proliferation of PC3 cells ( IC(50)= 250.21\u2009\u03bcg/mL ) when compared with a chemotherapeutic anticancer drug ( Toxol ) ( Vesper Pharmaceuticals ) with increased nucleosome production from apoptotic cells .\n\nThe Punica granatum L. var. spinosa extract attenuated the human prostate cell proliferation in vitro possibly by inducing apoptosis .\n\nThe Punica granatum L. var. spinosa is likely to be valuable for the treatment of some forms of human prostate cell line .", "output": "Resisting cell death" }, { "input": "OBJECTIVE The signalling molecule protein kinase B ( Akt ) modulates many cellular processes .\n\nPhosphatidylinositol 3-kinase ( PI3K)/Akt signalling pathways play important roles in tumour angiogenesis .\n\nThe aim of this study was to determine the role of phosphorylated Akt ( pAkt ) in angiogenesis and its correlation with vascular endothelial growth factor ( VEGF)-A in gastric adenocarcinoma .\n\nMETHODS Tumour tissue and matched healthy gastric mucosa were obtained from patients undergoing surgical resection of gastric adenocarcinoma .\n\nAkt and pAkt were detected via Western blotting .\n\nVEGF-A , pAkt and CD34 were examined by immunohistochemistry .\n\nRESULTS Akt and pAkt protein levels were significantly higher in gastric cancer tissue than in normal tissue ( n = 48 patients ) .\n\nPositive VEGF-A immunostaining was significantly associated with pAkt immunostaining .\n\nMicrovessel density was correlated with both VEGF-A and pAkt positivity .\n\nCONCLUSIONS Phosphorylated Akt and VEGF-A are involved in angiogenesis of gastric adenocarcinoma , and Akt activation may contribute to angiogenesis via VEGF-A upregulation .\n\nThe PI3K/Akt/VEGF signalling pathway may be involved in gastric adeno carcinoma .", "output": "Inducing angiogenesis, Sustaining proliferative signaling" }, { "input": "This study aimed to observe the effects of octreotide ( OCT ) on cisplatin resistance reversal of cancer cells in vitro and in nude mice in vivo .\n\nMTT method and flow cytometry were used to investigate the effect of cisplatin , OCT or the combination of these two compounds on the proliferation and apoptosis of SKOV3-DDP cells .\n\nThe size and weight of xenograft tumors from the nude mice model were measured .\n\nReal-time PCR was used to detect the mRNA expression of SSTR2 , MDR1 , MRP2 , GST-pi and EGFR in SKOV3/DDP cells following the different treatment .\n\nAt the concentration of 2.5-20 g/ml , OCT significantly reduced IC50 ( p < 0.05 ) and promoted apoptosis ( p < 0.05 ) of SKOV3-DDP cells ' response to cisplatin .\n\nUnchanged expression was found in SSTR2 on the SKOV3/DDP cell in vitro after OCT treatment , but increased expression in vivo ( p < 0.05 ) .\n\nOCT increased GST-pi expression ( p < 0.05 ) and reduced MRP2 and EGFR expression ( p < 0.05 ) in a dose-dependent manner .\n\nThe similar results were obtained in mice in vivo experiment , except the reduced expression of GST-pi .\n\nIt is suggested that OCT could inhibit ovarian cancer proliferation and promote apoptosis , via the cell surface SSTR2 , and reverse cisplatin resistance through inhibition of MRP2 , EGFR , and even GST-pi expressions .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "UVB is a major cause of nonmelanoma skin cancer in humans .\n\nPhotochemoprevention represents an important strategy in protecting the skin against the detrimental effects of ultraviolet B ( UVB ) .\n\nWe investigated the activity of Calluna vulgaris ( Cv ) delivered via a hydrogel on 3 main pathways ( oxidative stress , inflammation , DNA damage ) on skin exposed to multiple doses of UVB in SKH-1 mice .\n\nFifty female mice were divided randomly into 5 groups : control , vehicle , UVB irradiated , Cv + UVB irradiated , and Cv + vehicle + UVB irradiated .\n\nThe extract was applied topically on the skin in a dose of 4 mg polyphenols/cm2 30 minutes before each UVB ( 240 mJ/cm2 ) exposure over 10 consecutive days .\n\nMalondialdehyde , reduced glutathione , tumor necrosis factor-\u03b1 , interleukin-6 , cyclobutane pyrimidine dimer ( CPD ) levels , sunburn cell formation and epidermal thickness , and the number of epidermal cell layers in skin were evaluated 24 hours after the last treatment .\n\nUVB increased cytokine levels ( P < 0.001 ) , formation of CPDs ( P < 0.001 ) and sunburn cells ( P < 0.001 ) , and the epidermal thickness and number of epidermal cell layers ( P < 0.001 ) compared with the control group .\n\nThe topical application of Cv protected the skin against inflammation and DNA damage , as shown by a decreased number of CPDs ( P < 0.001 ) and sunburn cells ( P < 0.001 ) .\n\nThe administration of Cv via hydrogel may be a viable method for chemoprevention. .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "Epithelial-mesenchymal transition ( EMT ) is a critical process providing tumor cells with the ability to migrate and escape from the primary tumor and metastasize to distant sites .\n\nRecently , EMT was shown to be associated with the cancer stem cell ( CSC ) phenotype in breast cancer .\n\nSnail is a transcription factor that mediates EMT in a number of tumor types , including colorectal cancer ( CRC ) .\n\nOur study was done to determine the role of Snail in mediating EMT and CSC function in CRC .\n\nHuman CRC specimens were stained for Snail expression , and human CRC cell lines were transduced with a retroviral Snail construct or vector control .\n\nCell proliferation and chemosensitivity to oxaliplatin of the infected cells were determined by the MTT ( colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ) assay .\n\nMigration and invasion were determined in vitro using modified Boyden chamber assays .\n\nEMT and putative CSC markers were analyzed using Western blotting .\n\nIntravenous injection of tumor cells was done to evaluate their metastatic potential in mice .\n\nSnail was overexpressed in human CRC surgical specimens .\n\nThis overexpression induced EMT and a CSC-like phenotype in human CRC cells and enhanced cell migration and invasion ( P < 0.002 vs. control ) .\n\nSnail overexpression also led to an increase in metastasis formation in vivo ( P < 0.002 vs. control ) .\n\nFurthermore , the Snail-overexpressing CRC cells were more chemoresistant to oxaliplatin than control cells .\n\nIncreased Snail expression induces EMT and the CSC-like phenotype in CRC cells , which enhance cancer cell invasion and chemoresistance .\n\nThus , Snail is a potential therapeutic target in metastatic CRC .", "output": "Activating invasion and metastasis" }, { "input": "The purpose of the study was to evaluate Sorafenib ( BAY 43-9006 ) derived receptor tyrosine kinase inhibition on tumor progression in murine islet cell tumors .\n\nSorafenib is considered to be a potent inhibitor of tumor angiogenesis and neovascularization in various solid tumors .\n\nRip1Tag2 mice were treated in two different groups according to the model of tumor progression : the early treatment group received vehicle or Sorafenib from 10 to 14 weeks of age and the late treatment group from week 12 until death .\n\nTumor surface , tumor cell proliferation , and apoptosis were measured in both treatment groups to assess the in vivo effects of Sorafenib .\n\nSurvival was recorded for the late treatment group .\n\nIn the early treatment group Sorafenib led to a dramatic decrease in tumor volume compared to the control group .\n\nApoptosis was significantly augmented and cell proliferation was inhibited .\n\nAs a single therapy Sorafenib significantly improved survival in the late treatment group .\n\nConclusion .\n\nSorafenib may provide a new paradigm for the therapy of islet cell tumors .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "Cellular redox changes have emerged as a pivotal and proximal event in cancer .\n\nPKI 166 is used to determine the effects of redox sensitive inhibition of EGFR , metastasis and apoptosis in epidermoid carcinoma .\n\nCytotoxicity study of PKI 166 ( IC50 1.0 microM ) treated A431 cells were performed by MTT assay for 48 and 72 hrs .\n\nMorphological analysis of PKI 166 treated A431 cells for 48 hrs. revealed the cell shrinkage , loss of filopodia and lamellipodia by phase contrast and SEM images in dose dependent manner .\n\nIt has cytotoxic effects through inhibiting cellular proliferation , leads to the induction of apoptosis , as increased fraction of sub-G1 phase of the cell cycle , chromatin condensation and DNA ladder .\n\nIt inhibited cyclin-D1 and cyclin-E expression and induced p53 , p21 expression in dose dependent manner .\n\nConsequently , an imbalance of Bax/Bcl-2 ratio triggered caspase cascade and subsequent cleavage of PARP , thereby shifting the balance in favour of apoptosis .\n\nPKI 166 treatment actively stimulated reactive oxygen species ( ROS ) and mitochondrial membrane depolarization .\n\nIt inhibited some metastatic properties of A431 cells supressing colony formation by soft agar assay and inhibition of MMP 9 activity by gelatin zymography and western blot analysis .\n\nPKI 166 inhibited growth factor induced phosphorylation of EGFR , Akt , MAPK , JNK and colony formation in A431 cells .\n\nThus the inhibition of proliferation was associated with redox regulation of the caspase cascade , EGFR , Akt/PI3K , MAPK/ ERK and JNK pathway .\n\nOn the other hand , increased antioxidant activity leads to decreased ROS generation inhibit the anti-proliferative and apoptotic properties of PKI 166 in A431 cells .\n\nThese observations indicated PKI 166 induced redox signalling dependent inhibition of cell proliferation , metastatic properties and induction of apoptotic potential in epidermoid carcinoma .", "output": "Activating invasion and metastasis, Sustaining proliferative signaling, Evading growth suppressors, Tumor promoting inflammation, Resisting cell death" }, { "input": "The Hippo signaling pathway inhibits cell growth and regulates organ size through a kinase cascade that leads to the phosphorylation and nuclear exclusion of the growth-promoting transcriptional coactivator Yes-associated protein ( YAP)/Yorkie .\n\nIt mediates contact inhibition of cell growth downstream of cadherin adhesion molecules and other cell surface proteins .\n\nContact inhibition is often antagonized by mitogenic growth factor signaling .\n\nWe report an important mechanism for this antagonism , inhibition of Hippo pathway signaling by mitogenic growth factors .\n\nEGF treatment of immortalized mammary cells triggers the rapid translocation of YAP into the nucleus along with YAP dephosphorylation , both of which depend on Lats , the terminal kinase in the Hippo pathway .\n\nA small-molecule inhibitor screen of downstream effector pathways shows that EGF receptor inhibits the Hippo pathway through activation of PI3-kinase ( PI3K ) and phosphoinositide-dependent kinase ( PDK1 ) , but independent of AKT activity .\n\nThe PI3K-PDK1 pathway also mediates YAP nuclear translocation downstream of lysophosphatidic acid and serum as a result of constitutive oncogenic activation of PI3K .\n\nPDK1 associates with the core Hippo pathway-kinase complex through the scaffold protein Salvador .\n\nThe entire Hippo core complex dissociates in response to EGF signaling in a PI3K-PDK1-dependent manner , leading to inactivation of Lats , dephosphorylation of YAP , and YAP nuclear accumulation and transcriptional activation of its target gene , CTGF .\n\nThese findings show that an important activity of mitogenic signaling pathways is to inactivate the growth-inhibitory Hippo pathway and provide a mechanism for antagonism between contact inhibition and growth factor action .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "OBJECTIVE To observe the anti-tumor recurrent and metastatic efficacy of Ru'ai Shuhou Recipe ( RSR ) on HER2 positive breast cancer , to evaluate the effects of RSR on the expressions of matrix metalloproteinases ( MMPs ) and the tissue inhibitor of metalloproteinases ( TIMPs ) in the recurrence and metastasis of HER2 positive breast cancer , thus revealing its anti-tumor recurrent and metastatic mechanisms .\n\nMETHODS Selected were 30-week-old HER2/neu transgenic spontaneous breast cancer mice FVB/neu .\n\nThe primary tumor resection was carried out .\n\nAfter surgery they were randomly divided into the blank control group , the RSR group , the Herceptin group , and the combination group ( RSR + Herceptin group ) .\n\nThe treatment lasted for 4 months .\n\nThe inhibition rate of the recurrent tumor volume and the inhibition rate of the lung metastasis were evaluated .\n\nThe expressions of matrix metalloproteinase-2 ( MMP-2 ) , matrix metalloproteinase-9 ( MMP-9 ) , tissue inhibitor of metalloproteinase ( TIMP-1 ) , and TIMP-2 in the recurrent tumor tissue were detected using Western blot .\n\nRESULTS By the end of the treatment the average recurrent tumor volume was 11.11 +/- 8.71 cm3 in the blank control group and 5.56 +/- 5.55 cm3 of the RSR group , showing statistical difference between the two groups ( P = 0.037 ) .\n\nThe average lung metastatic nodule was 16 in the blank control group and 10 in the RSR group .\n\nThe inhibition rate of lung metastasis was 37. 85% in the RSR group , but with no statistical significance .\n\nThe expression level of activated MMP-2 in the RSR group was down-regulated when compared with the blank control group , the Herceptin group , and the combination group ( P < 0.05 ) .\n\nThe expression of MMP-9 of the RSR group , the Herceptin group , and the combination group was significantly down-regulated when compared with the blank control group ( P < 0.05 ) .\n\nThe expression of MMP-9 of the RSR group and the combination group was further down-regulated when compared with the Herceptin group ( P < 0.05 ) .\n\nThe expressions of both TIMP-1 and TIMP-2 of the RSR group , the Herceptin group , and the combination group were all up-regulated when compared with the blank control group ( P < 0.05 ) .\n\nThe increased expression of TIMP-1 was more significantly in the RSR group and the combination group when compared with the Herceptin group ( P < 0.05 ) .\n\nIt was higher in the combination group than in the RSR group ( P < 0.05 ) .\n\nCONCLUSIONS RSR could inhibit the tumor recurrence of FVB/neu mice .\n\nIt could reduce the degradation of extracellular matrix and increase the protective effects of extracellular matrix .\n\nIt might achieve its anti-tumor effect through effecting the invasive and metastatic capabilities of breast tumor cells .", "output": "Activating invasion and metastasis" }, { "input": "Bcl-2/E1B 19-kDa interacting protein 3 ( BNIP3 ) is a proapoptotic protein whose expression level is often low in colorectal cancer ( CRC ) cells due to the BNIP3 gene promoter DNA methylation by DNA methyltransferase ( DNMT ) .\n\nIt is known that chemotherapy and radiotherapy suppress CRC through inducing tumor apoptosis .\n\nHowever , the molecular mechanisms underlying chemotherapy and radiotherapy-induced apoptosis of CRC cells are not well defined .\n\nIn this study , we observed that the expression level of BNIP3 in colon cancer cells was significantly increased by treatment with therapeutic agents and radiation in vitro .\n\nThe BNIP3 protein level in CRC tissues from patients who received preoperative concurrent chemotherapy was significantly higher than in those who received surgery alone .\n\nFurthermore , treatment with chemotherapeutic agents and radiation significantly decreased the DNMT1 expression level and enzymatic activity .\n\nBoth expression level and activity of DNMT1 were inversely correlated with the expression level of BNIP3 in colon carcinoma cells after treatment with chemotherapeutic agents and radiation .\n\nConsistent with increased BNIP3 expression , chemotherapeutic agents and radiation induced colon carcinoma cell apoptosis in a dose-dependent manner .\n\nBased on these observations , we conclude that chemotherapy and radiotherapy inhibit DNMT1 expression to upregulate BNIP3 expression to promote CRC cell apoptosis .\n\nAnd , BNIP3 may play a role in the caspase-dependent apoptosis pathways , mainly during treatment with chemotherapy and radiotherapy .", "output": "Resisting cell death" }, { "input": "The new concept of keeping primary tumor under control in situ to suppress distant foci sheds light on the novel treatment of metastatic tumor .\n\nHyperthermia is considered as one of the means for controlling tumor growth .\n\nIn this study , a novel thermal modality was built to introduce hyperthermia effect on tumor to suppress its growth and progression using 4T1 murine mammary carcinoma , a common animal model of metastatic breast cancer .\n\nA mildly raised temperature ( i.e.39\ufffdC ) was imposed on the skin surface of the implanted tumor using a thermal heating pad .\n\nPeriodic heating ( 12 hours per day ) was carried out for 3 days , 7 days , 14 days , and 21 days , respectively .\n\nThe tumor growth rate was found significantly decreased in comparison to the control without hyperthermia .\n\nBiological evidences associated with tumor angiogenesis and metastasis were examined using histological analyses .\n\nAccordingly , the effect of mild hyperthermia on immune cell infiltration into tumors was also investigated .\n\nIt was demonstrated that a delayed tumor growth and malignancy progression was achieved by mediating tumor cell apoptosis , vascular injury , degrading metastasis potential and as well as inhibiting the immunosuppressive cell myeloid derived suppressor cells ( MDSCs ) recruitment .\n\nFurther mechanistic studies will be performed to explore the quantitative relationship between tumor progression and thermal dose in the near future .", "output": "Activating invasion and metastasis, Avoiding immune destruction, Resisting cell death" }, { "input": "UNLABELLED BACKGROUND Cell lines are very useful for clinical and basic research .\n\nThus far , only 11 reports have documented the characteristics of ovarian endometrioid adenocarcinoma cell lines in the literature .\n\nDue to the scarcity of information , the establishment of an ovarian malignant tumor cell line with distinctive characteristics is particularly important to study this disease .\n\nThus , this study was undertaken to establish and characterize a new human endometrioid adenocarcinoma cell line of the ovary .\n\nMETHODS The cell line NOMH-1 was established from an ovarian tumor of a 44-year-old woman .\n\nFeatures of the cell line studied included morphology , chromosome analysis , heterotransplantation , tumor markers , and chemosensitivity .\n\nRESULTS This cell line has been growing well for 232 months and subcultured more than 50 times .\n\nMonolayer cultured cells were polygonal in shape , showing a pavement-like arrangement and a tendency to stack without contact inhibition .\n\nThey exhibited a human karyotype with a modal chromosomal number in the hypertriploid range .\n\nThe cells could be transplanted into the subcutis of nude mice and produced tumors resembling the original tumor .\n\nNOMH-1 cells expressed both CEA and CA19-9 which were identified immunohistochemically in the original tumor and the heterotransplanted tumor .\n\nThe cells were sensitive to paclitaxel , an agent commonly used in the treatment of gynecological cancers .\n\nCONCLUSIONS NOMH-1 is the first ovarian endometrioid adenocarcinoma cell line in which CEA and CA19-9 expression have been defined .\n\nThis newly established cell line should be useful for investigating the characteristics of ovarian endometrioid adenocarcinoma .", "output": "Evading growth suppressors" }, { "input": "The effects of genomic changes in hepatitis B virus ( HBV ) on the occurrence of hepatocellular carcinoma ( HCC ) are still unclear , especially in relation to the genotype of HBV .\n\nIn this study , we examined the effects of genomic changes in HBV of genotype C2 on the development of HCC .\n\nA total of 318 patients with HBV-associated HCC and 234 patients with chronic hepatitis B ( CHB ) were studied .\n\nAll of HCC cases were diagnosed histologically and treated with surgical resection .\n\nThe whole of the X , S , basal core promoter ( BCP ) and precore regions of the viral genome from sera or liver tissues were sequenced .\n\nAll subjects had HBV of genotype C2 .\n\nThe prevalence of the T1653 mutation in the X region and the A1896 mutation in the precore region of HBV was significantly higher in the HCC group than in the control CHB group ( 22% vs 11% , P = 0.003 ; 50% vs 23% , P < 0.001 , respectively ) .\n\nMoreover , the T1762/A1764 mutations in the BCP region in combination with either T1653 or A1896 were more common in the HCC compared with the CHB group ( BCP+X1653 : 18% vs 11% , P = 0.05 ; BCP+PC , 40% vs 15% , P < 0.001 , respectively ) .\n\nIn multivariate analysis , T1653 and A1896 were revealed to be independent risk factors for HCC development .\n\nG1896A in the precore region and C1653T mutation in the X region of genotype C2 HBV are important risk factors for HCC development .\n\nAlso , the A1762T/G1764A double mutation may act in synergy with C1653T to increase the risk of HCC in patients chronically infected with HBV genotype C2 .", "output": "Genomic instability and mutation" }, { "input": "OBJECTIVE This study describes a new method used in the clinical laboratory at Hospital Israelita Albert Einstein to detect mutations in exons 8 and 17 of the KIT gene in patients with acute myeloid leukemia .\n\nMETHODS Genomic DNA extraction was performed on 54 samples of peripheral blood or bone marrow from patients with acute myeloid leukemia .\n\nThe extracted DNA was amplified by polymerase chain reaction and sequenced , and the fragments were analyzed .\n\nRESULTS Within the analyzed samples , we detected four mutations in exon 8 , two mutations in exon 17 , and mutations or a double mutation in one sample .\n\nCONCLUSION The tests detecting mutations in exon 8 and 17 on the KIT gene were successfully standardized .\n\nThe test is now included among the routine diagnostics employed for patients at Hospital Israelita Albert Einstein clinical laboratory .", "output": "Genomic instability and mutation" }, { "input": "In the present study , post-treatment effects of dietary turmeric on markers related to apoptosis , cell proliferation , and inflammation in 7,12-dimethylbenz(a)anthracene ( DMBA)-induced hamster buccal pouch ( HBP ) tumors were investigated .\n\nTumors were induced by applying 0.5% DMBA topically to the HBP three times per week for 12 weeks .\n\nAfter tumor development , half of the animals continued on the control diet and the other half were shifted to a 1% turmeric diet for 4 weeks .\n\nTo rule out DMBA discontinuation as a cause of inhibition in tumor growth , DMBA treatment was continued during dietary exposure of turmeric in another set of animals until the end of the experiment .\n\nThe turmeric diet inhibited tumor growth in animals with or without DMBA carcinogen treatment compared to the animals on the control diet .\n\nWhen compared to hamsters bearing tumors that remained on the control diet , the buccal pouches of hamsters bearing tumors receiving turmeric showed the following results : ( 1 ) decreased cell proliferation ( diminished PCNA , cyclin D1 , and Bcl-2 ) and PCNA labelling index , ( 2 ) enhanced apoptosis ( increased Bax , caspase-3 , caspase-9 , and cytochrome c , and decreased survivin ) and apoptotic index , ( 3 ) decreased inflammation ( decreased Cox-2 ) , and ( 4 ) decreased MAPK activation ( p-ERK and p-p38 ) .\n\nThese data indicate that tumor growth decreased due to the modulation of cellular pathways associated with cell proliferation and apoptosis .", "output": "Tumor promoting inflammation, Sustaining proliferative signaling, Resisting cell death" }, { "input": "OBJECTIVE To sort and identify side population ( SP ) cancer stem cells ( CSC ) in human prostate cancer ( PCa ) cell lines .\n\nMETHODS Stem-like cells were isolated from five PCa cell lines Du145 , IA8 , LNCaP , TSU-Pr and PC-3 using FACS based on CD133+ CD44+ immunophenotype and SP in Hoechst staining .\n\nThe in vitro growth pattern and tumorigenicity of SP stem cells were verified by soft agar colony-formation trial .\n\nLNCaP/SP cells were selected for further identification of stem cell properties using immunostaining , proliferation and invasion assay .\n\nEventually , tumorigenicity and metastasis ability of LNCaP/SP were confirmed by xenograft experiments .\n\nRESULTS The percentages of CSCs of the CD133 CD44 + immunophenotype were extremely low in the five PCa cell lines .\n\nOn the contrary , the percentages of the isolated SP cells were significantly higher in Du145 ( [ 0.15 +/- 0.02]% ) , IA8 ( [ 0.60 +/- 0.07 ]% ) , LNCaP ( [ 0.8 +/- 0.1]% ) and TSU-PrL ( [ 2.0 +/- 0.4]% ) , but none was detected in PC-3 .\n\nBesides , IA8/SP , LNCaP/SP and TSU-PrL/SP cells showed a significantly greater colony-forming efficiency than non-side population ( NSP ) cells ( P < 0.05 ) .\n\nCompared with LNCaP/NSP cells , LNCaP/SP cells exhibited high expressions of integrin alpha2 , Nanog , CD44 , OCT4 and ABCG2 , remarkably enhanced invasive and proliferative potentials in vitro , and markedly increased tumorigenicity and metastasis ( P < 0.01 ) .\n\nCONCLUSION SP sorting is more suitable than CD133+ CD44+ selection for enriching CSCs from PCa cell lines , and LNCaP/ SP represents a typical CSC population .", "output": "Activating invasion and metastasis" }, { "input": "BACKGROUND DNA-damaging compounds ( e.g. , alkylating agents , cytotoxic antibiotics and DNA topoisomerase poisons ) are the most widely used anticancer drugs .\n\nThe inability of tumor cells to properly repair some types of DNA damage may explain why specific DNA-damaging drugs can selectively kill tumor cells .\n\nPhenylglyoxal is a dicarbonyl compound known to react with guanidine groups such as that of the DNA base guanine , therefore suggesting that phenylglyoxal could induce DNA damage and have anticancer activity .\n\nMETHODS Cellular DNA damage was measured by the alkaline comet assay and the \\u03b3H2AX focus assay .\n\nFormation of topoisomerase I- and topoisomerase II-DNA complexes was assessed by the TARDIS assay , an immunofluorescence technique that employs specific antibodies to DNA topo I or topo II to detect the protein covalently bound to the DNA in individual cells .\n\nCell growth inhibition and cytotoxicity were determined by XTT , MTT and clonogenic assays .\n\nApoptosis was assessed by the Annexin V flow cytometry assay .\n\nRESULTS Phenylglyoxal induced cellular DNA damage and formation of high levels of topoisomerase I- and topoisomerase II-DNA complexes in cells .\n\nThese topoisomerase-DNA complexes were abolished by catalase pretreatment and correlated well with the induction of apoptosis .\n\nPhenylglyoxal-induced cell death was partially prevented by catalase pretreatment and was higher in lung cancer cells ( A549 ) than in normal lung fibroblasts ( MRC5 ) .\n\nMammalian cell lines defective in nucleotide excision repair ( NER ) , homologous recombination ( HR ) and non-homologous end joining ( NHEJ ) were more sensitive to phenylglyoxal than parental cells ; this suggests that phenylglyoxal may induce bulky distortions in the shape of the DNA double helix ( which are repaired by the NER pathway ) and DNA double-strand breaks ( which are repaired by HR and NHEJ ) .\n\nCONCLUSION This report shows that phenylglyoxal is a new DNA-damaging agent with anticancer activity , and suggests that tumor cells with defects in NER , HR and NHEJ may be hypersensitive to the cytotoxic activity of phenylglyoxal .", "output": "Genomic instability and mutation, Resisting cell death" }, { "input": "We have previously shown that the antiprogestin and antiglucocorticoid mifepristone inhibits the growth of ovarian cancer cells .\n\nIn this work , we hypothesized that cellular stress caused by mifepristone is limited to cytostasis and that cell killing is avoided as a consequence of the persistent activity of the PI3K/Akt survival pathway.To investigate the role of this pathway in mifepristone-induced growth inhibition , human ovarian cancer cells of various histological subtypes and genetic backgrounds were exposed to cytostatic doses of mifepristone in the presence or absence of the PI3K inhibitor , LY294002 .\n\nThe activation of Akt in ovarian cancer cells , as marked by its phosphorylation on Ser473 , was not modified by cytostatic concentrations of mifepristone , but it was blocked upon treatment with LY294002 .\n\nThe combination mifepristone/LY294002 , but not the individual drugs , killed ovarian cancer cells via apoptosis , as attested by genomic DNA fragmentation and cleavage of caspase-3 , and the concomitant down-regulation of anti-apoptotic proteins Bcl-2 and XIAP .\n\nFrom a pharmacological standpoint , when assessing cell growth inhibition using a median-dose analysis algorithm , the interaction between mifepristone and LY294002 was synergistic .\n\nThe lethality caused by the combination mifepristone/LY294004 in two dimensional cell cultures was recapitulated in organized , tri-dimensional spheroids .\n\nThis study demonstrates that mifepristone and LY294002 , when used individually , cause cell growth arrest , yet when combined , they cause lethality .", "output": "Resisting cell death" }, { "input": "HM781-36B is an orally administered pan-human epidermal growth factor receptor ( HER ) inhibitor .\n\nTo explore the role of pan-HER inhibitor in breast cancer , we investigated the antitumor effect and mechanisms of HM781-36B in breast cancer cell lines .\n\nSix breast cancer cell lines ( BT474 , MDA-MB-453 , SK-BR-3 , T47D , MCF-7 , and MDA-MB-231 ) were tested .\n\nThe growth inhibitory effect was assessed using the tetrazolium bromide [ 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl-tetrazolium bromide ] assay .\n\nThe cell cycle at various concentrations of HM781-36B was analyzed by flow cytometry , and analysis of downstream molecules was performed by western blot analysis .\n\nInteraction of HM781-36B with cytotoxic chemotherapeutic agents was analyzed by combination index using CalcuSyn .\n\nThe HER2-amplified cells ( SK-BR-3 , BT474 , and MDA-MB-453 ) were sensitive to HM781-36B ( IC50=0.001 \u03bcmol/l , 0.0012 \u03bcmol/l , and 0.0095 \u03bcmol/l , respectively ) .\n\nHM781-36B induced G1 arrest and resulted in apoptosis .\n\nIt reduced the level of p-HER2 , p-AKT , p-ERK , and p-STAT3 .\n\nHM781-36B combined with 5-fluorouracil , cisplatin , paclitaxel , or gemcitabine showed a synergistic inhibitory effect on the HER2-amplified and on some of the HER2-nonamplified breast cancer cells .\n\nHM781-36B could be a promising treatment for HER2-amplified breast cancer as a single agent or in combination with cytotoxic agents and can be a candidate for treatment of HER2-nonamplified breast cancer in combination with cytotoxic agents .", "output": "Sustaining proliferative signaling, Resisting cell death" }, { "input": "Disruption of contact inhibition and serum afflux that occur after a tissue injury activate cell cycle , which then stops when confluence is reached again .\n\nAlthough the events involved in cell cycle entry have been widely documented , those managing cell cycle exit have remained so far ill defined .\n\nWe have identified that the final stage of wound closure is preceded in keratinocytes by a strong accumulation of miR-483-3p , which acts as a mandatory signal triggering cell cycle arrest when confluence is reached .\n\nBlocking miR-483-3p accumulation strongly delays cell cycle exit , maintains cells into a proliferative state and retards their differentiation program .\n\nUsing two models of cell cycle synchronization ( i.e. mechanical injury and serum addition ) , we show that an ectopic upregulation of miR-483-3p blocks cell cycle progression in early G1 phase .\n\nThis arrest results from a direct targeting of the CDC25A phosphatase by miR-483-3p , which can be impeded using an anti-miRNA against miR-483-3p or a protector that blocks the complex formation between miR-483-3p and the 3'-untranslated region ( UTR ) of CDC25A transcript .\n\nWe show that the miRNA-induced silencing of CDC25A increases the tyrosine phosphorylation status of CDK4/6 cyclin-dependent kinases which , in turn , abolishes CDK4/6 capacity to associate with D-type cyclins .\n\nThis prevents CDK4/6 kinases ' activation , impairs downstream events such as cyclin E stimulation and sequesters cells in early G1 .\n\nWe propose this new regulatory process of cyclin-CDK association as a general mechanism coupling miRNA-mediated CDC25A invalidation to CDK post-transcriptional modifications and cell cycle control .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "A new cell line of human ovarian clear cell adenocarcinoma ( CCC ) , TU-OC-1 , was established and characterized .\n\nThe cells showed a polygonal-shaped morphology and grew in monolayers without contact inhibition and were arranged like a jigsaw puzzle .\n\nThe chromosome numbers ranged from 64 to 90 .\n\nA low rate of proliferation was observed , similar to other CCC cell lines tested ( OVTOKO , RMG-I , and OVAS ) , and the doubling time was 38.4h .\n\nThe respective IC50 values of cisplatin and paclitaxel were 12.2\u03bcM and 58.3nM .\n\nMutational analysis revealed that TU-OC-1 cells harbored a PIK3CA mutation at codon 542 ( E542K ) in exon 9 , which is a mutation hot spot on this gene .\n\nWe observed that phosphorylated Akt protein was overexpressed in TU-OC-1 cells by western blot analysis .\n\nHeterotransplantation to nude mice produced tumors that reflected the original .\n\nThis cell line may be useful to study the chemoresistant mechanisms of CCC and contribute to novel treatment strategies .", "output": "Genomic instability and mutation" }, { "input": "Autophagy is a catabolic process by which a cell degrades its intracellular materials to replenish itself .\n\nInduction of autophagy under various cellular stress stimuli can lead to either cell survival or cell death via apoptotic and/or autophagic ( nonapoptotic ) pathways .\n\nThe NSAID sulindac sulfide induces apoptosis in colon cancer cells .\n\nHere , we show that inhibition of autophagy under serum-deprived conditions resulted in significant reductions of sulindac sulfide-induced apoptosis in HT-29 colon cancer cells .\n\nIn contrast , inhibition of autophagy under conditions where serum is available significantly increased sulindac sulfide-induced apoptosis in HT-29 cells .\n\nWe previously showed that the apoptosis inhibitor , survivin , plays a role in regulating NSAID-induced apoptosis and autophagic cell death .\n\nHere , we show that survivin protein half-life is increased in the presence of autophagy inhibitors under serum-deprived conditions , but not under conditions when serum is available .\n\nThus , the increased levels of survivin may be a factor contributing to inhibition of sulindac sulfide-induced apoptosis under serum-deprived conditions .\n\nThese results suggest that whether a cell lives or dies due to autophagy induction depends on the balance of factors that regulate both autophagic and apoptotic processes .", "output": "Resisting cell death" }, { "input": "We recently reported that chronic myeloid leukaemia ( CML ) patients harbour high levels of STAT5 when they progress to advanced phases of disease .\n\nAdvanced disease is characterized by an increased incidence of BCR-ABL1 mutations .\n\nWe now describe a highly significant correlation between STAT5 expression and the incidence of BCR-ABL1 mutations in primary CML .\n\nForced expression of STAT5 in murine BCR-ABL1 transformed cells sufficed to enhance the production of reactive oxygen species ( ROS ) and to trigger DNA damage .\n\nSTAT5-mediated ROS production is independent of JAK2 but requires concomitant BCR-ABL1 signalling as forced STAT5 expression in untransformed BCR-ABL1 negative cells has no impact on ROS levels .\n\nOnly within the context of a BCR-ABL1 positive cell does STAT5 transcriptionally regulate a target gene or set of genes that causes the enhanced ROS production .\n\nOur study suggests the existence of a feed-forward loop accelerating disease progression , in which BCR-ABL1 enhances its own mutation rate in a STAT5-ROS dependent manner .\n\nThis model explains the increased occurrence of inhibitor-resistant BCR-ABL1 mutations in advanced disease stages driven and characterized by high STAT5 expression .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "Transforming growth factor beta ( TGF-\u03b2 ) signaling has been implicated in driving tumor progression and metastasis by inducing stem cell-like features in some human cancer cell lines .\n\nIn this study , we have utilized a novel murine cell line NMuMG-ST , which acquired cancer stem cell ( CSC ) phenotypes during spontaneous transformation of the untransformed murine mammary cell line NMuMG , to investigate the role of autocrine TGF-\u03b2 signaling in regulating their survival , metastatic ability , and the maintenance of cancer stem cell characteristics .\n\nWe have retrovirally transduced a dominant-negative TGF-\u03b2 type II receptor ( DNRII ) into the NMuMG-ST cell to abrogate autocrine TGF-\u03b2 signaling .\n\nThe expression of DNRII reduced TGF-\u03b2 sensitivity of the NMuMG-ST cells in various cell-based assays .\n\nThe blockade of autocrine TGF-\u03b2 signaling reduced the ability of the cell to grow anchorage-independently and to resist serum deprivation-induced apoptosis .\n\nThese phenotypes were associated with reduced levels of active and phosphorylated AKT and ERK , and Gli1 expression suggesting that these pathways contribute to the growth and survival of this model system .\n\nMore interestingly , the abrogation of autocrine TGF-\u03b2 signaling also led to the attenuation of several features associated with mammary stem cells including epithelial-mesenchymal transition , mammosphere formation , and expression of stem cell markers .\n\nWhen xenografted in athymic nude mice , the DNRII cells were also found to undergo apoptosis and induced significantly lower lung metastasis burden than the control cells even though they formed similar size of xenograft tumors .\n\nThus , our results indicate that autocrine TGF-\u03b2 signaling is involved in the maintenance and survival of stem-like cell population resulting in the enhanced metastatic ability of the murine breast cancer cells .", "output": "Activating invasion and metastasis, Resisting cell death" }, { "input": "Systemic oxidative stress is associated with a wide range of pathological conditions .\n\nOxidative DNA damage is frequently measured in circulating lymphocytes .\n\nMitochondrial DNA ( mtDNA ) is known to be more sensitive to oxidative damage than nuclear DNA but is rarely used for direct measurement of DNA damage in clinical studies .\n\nBased on the supercoiling-sensitive real-time PCR method , we propose a new approach for the noninvasive monitoring of systemic oxidative stress by quantifying the mtDNA structural damage and copy number change in isolated lymphocytes in a single test .\n\nWe show that lymphocytes have significantly less mtDNA content and relatively lower baseline levels of damage than cancer cell lines .\n\nIn an ex vivo challenge experiment , we demonstrate , for the first time , that exogenous H2O2 induces a significant increase in mtDNA damage in lymphocytes from healthy individuals , but no repair activity is observed after 1 h recovery .\n\nWe further demonstrate that whole blood may serve as a convenient alternative to the isolated lymphocytes in mtDNA analysis .\n\nThus , the blood analysis with the multiple mtDNA end-points proposed in the current study may provide a simple and sensitive test to interrogate the nature and extent of systemic oxidative stress for a broad spectrum of clinical investigations .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "Metastases in the sellar region are rare and are frequently found incidentally or in necropsies .\n\nOnly 7% are reported to be symptomatic .\n\nDiabetes insipidus , anterior pituitary dysfunction , visual field defects , headache/pain and ophthalmoplegia are the most commonly reported symptoms .\n\nWe present the cases of two male patients with a small-cell lung carcinoma whose first clinical symptoms were due to pituitary metastasis .\n\nOne case presented with symptoms of cavernous sinus invasion and panhypopituitarism and the other case with diabetes insipidus .\n\nBoth patients had a rapid progression of their disease despite chemotherapy and died after a few months .\n\nPituitary metastases occur most commonly with breast cancer in women and lung cancer in men .\n\nThe presence of polyuria and polydipsia in an oncologic patient should alert the physician for diabetes insipidus and , if confirmed , an imaging procedure of the pituitary gland is mandatory .\n\nTreatment for these tumors is often multimodal and includes surgery , radiation therapy , chemotherapy and hormone replacement .\n\nAlthough surgical series have not shown any significant survival benefits given by tumor resection , the patient's quality of life may be improved .", "output": "Activating invasion and metastasis" }, { "input": "Within the family of protein kinase C ( PKC ) molecules , the novel isoform PRKCE ( PKC\u025b ) acts as a bona fide oncogene in in vitro and in vivo models of tumorigenesis .\n\nPrevious studies have reported expression of PKC\u025b in breast , prostate and lung tumors above that of normal adjacent tissue .\n\nData from the cancer genome atlas suggest increased copy number of PRKCE in triple negative breast cancer ( TNBC ) .\n\nWe find that overexpression of PKC\u025b in a non-tumorigenic breast epithelial cell line is sufficient to overcome contact inhibition and results in the formation of cellular foci .\n\nCorrespondingly , inhibition of PKC\u025b in a TNBC cell model results in growth defects in two-dimensional ( 2D ) and three-dimensional ( 3D ) culture conditions and orthotopic xenografts .\n\nUsing stable isotope labeling of amino acids in a cell culture phosphoproteomic approach , we find that CTNND1/p120ctn phosphorylation at serine 268 ( P-S268 ) occurs in a strictly PKC\u025b-dependent manner , and that loss of PKC\u025b signaling in TNBC cells leads to reversal of mesenchymal morphology and signaling .\n\nIn a model of Ras activation , inhibition of PKC\u025b is sufficient to block mesenchymal cell morphology .\n\nFinally , treatment with a PKC\u025b ATP mimetic inhibitor , PF-5263555 , recapitulates genetic loss of function experiments impairing p120ctn phosphorylation as well as compromising TNBC cell growth in vitro and in vivo .\n\nWe demonstrate PKC\u025b as a tractable therapeutic target for TNBC , where p120ctn phosphorylation may serve as a readout for monitoring patient response.Oncogene advance online publication , 1 April 2013 ; doi:10.1038/onc.2013.91 .", "output": "Evading growth suppressors" }, { "input": "Chronic alcohol ingestion increases hepatic cytochrome P450 2E1 ( CYP2E1 ) , which is associated with hepatocarcinogenesis .\n\nWe investigated whether treatment with chlormethiazole ( CMZ ) , a CYP2E1 inhibitor , protects against alcohol-associated hepatic carcinogenesis in rats .\n\nRats were fed either an ethanol liquid diet or a non-ethanol liquid diet , with or without CMZ for one and ten months .\n\nA single intraperitoneal injection of diethylnitrosamine ( DEN , 20 mg/kg ) was given to initiate hepatic carcinogenesis .\n\nCYP2E1 expression , inflammatory proteins , cell proliferation , protein-bound 4-HNE , etheno-DNA adducts , 8-hydroxy-2'-deoxyguanosine ( 8-OHdG ) , retinoid concentrations , and hepatic carcinogenesis were examined .\n\nEthanol feeding for 1 month with DEN resulted in significantly increased hepatic CYP2E1 levels and increased nuclear accumulation of NF-\\u03baB protein and TNF-\\u03b1 expression , which were associated with increased cyclin D1 expression and p-GST positive altered hepatic foci .\n\nAll of these changes induced by ethanol feeding were significantly inhibited by the one month CMZ treatment .\n\nAt 10-months of treatment , hepatocellular adenomas were detected in ethanol-fed rats only , but neither in control rats nor in animals receiving ethanol and CMZ .\n\nThe 8-OHdG formation was found to be significantly increased in ethanol fed animals and normalized with CMZ treatment .\n\nIn addition , alcohol-reduced hepatic retinol and retinoic acid concentrations were restored by CMZ treatment to normal levels in the rats at 10 months of treatment .\n\nThese data demonstrate that the inhibition of ethanol-induced CYP2E1 as a key pathogenic factor can counteract the tumor-promoting action of ethanol by decreasing TNF-\\u03b1 expression , NF-\\u03baB activation , and oxidative DNA damage as well as restoring normal hepatic levels of retinoic acid in DEN-treated rats .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "Cell migration plays important roles in natural processes involving embryonic development , inflammation , wound healing , cancer metastasis and angiogenesis .\n\nCell migration on various biomaterials is also believed to improve the rate of wound healing and implant therapies in the tissue-engineering field .\n\nThis study measured the distance traversed , or mileage , of mouse fibroblasts on a silk fibroin surface .\n\nFibroblasts on the fibroin surface moved with better progress during 24 h than cells on collagen or fibronectin surfaces .\n\nResults obtained by quantitative real-time reverse transcription-polymerase chain reaction ( qRT-PCR ) revealed that fibroblasts on the fibroin surface expressed transforming growth factor \\u03b2-induced protein ( TGFBI ) , which is an extracellular matrix ( ECM ) protein , stronger than on other surfaces in the early cell-culture stages .\n\nThese results demonstrate that the fibroin surface shows higher potential to enhance cell migration and the production of ECM than a collagen or fibronectin surface .", "output": "Activating invasion and metastasis" }, { "input": "BACKGROUND Angiogenesis is an essential process in cancer growth maintenance , and metastasis .\n\nAngiopoietin-2 promotes tumor angiogenesis by priming the vasculature and potentiating the effects of cytokines at the front of active neovascularization .\n\nEnhanced expression of angiopoietin-2 has been reported in lung cancer tissue .\n\nSurvivin is one of the inhibitors of apoptosis protein that has been shown to play a key role in cancer progression , and in tumor angiogenesis .\n\nAlso plays a key role in tumor cell resistance to anticancer agents and ionizing radiation .\n\nAIM To measure the serum levels of angiopoietin-2 and survivin as possible angiogenic factors in lung cancer patients with the assessment of their interrelationships and clinical significance .\n\nPATIENTS AND METHODS Patients with lung cancer as NSCLC ( n=70 ) and healthy volunteers ( n=10 ) were enrolled .\n\nSerum angiopoietin-2 and survivin concentrations were measured using enzyme-linked immunosorbent assay ( ELIZA ) .\n\nRESULTS Median serum angiopoietin-2 levels with lung cancer ( 2730pg/mL ) ranged from 1171 to 6541pg/mL was higher than the median of the control group ( 1795pg/mL ) ranged from 1076 to 2730/mL , p<0.001 .\n\nMedian serum survivin levels were also higher in patients with lung cancer ( 53.0pg/mL ) ranged from 39.3 to 96.3pg/mL than the median of the control group ( 48.8pg/mL ) ranged from 38.0 to 74.6pg/mL , but did not reach statistical significance p=0.206 .\n\nIn all patients with lung cancer , serum angiopoietin-2 was not significantly correlated with survivin ( r=0.073 , p=0.657 ) .\n\nNeither serum angiopoietin-2 nor survivin showed significant relation with the serum angiopoietin-2 or survivin levels depending on the cell types , stage progression , and metastasis among the patients with NSCLC .\n\nCONCLUSIONS Our study suggests that serum angiopoietin-2 is a useful marker for the diagnosis of NSCLC by ELIZA technique .", "output": "Inducing angiogenesis" }, { "input": "Understanding how arsenic exacts its diverse , global disease burden is hampered by a limited understanding of the particular biological pathways that are disrupted by arsenic and underlie pathogenesis .\n\nA reductionist view would predict that a small number of basic pathways are generally perturbed by arsenic , and manifest as diverse diseases .\n\nFollowing an initial observation that arsenite-exposed cells in culture acidify their media more rapidly than control cells , the report here shows that low level exposure to arsenite ( 75ppb ) is sufficient to induce aerobic glycolysis ( the Warburg effect ) as a generalized phenomenon in cultured human primary cells and cell lines .\n\nExpanded studies in one such cell line , the non-malignant pulmonary epithelial line , BEAS-2B , established that the arsenite-induced Warburg effect was associated with increased accumulation of intracellular and extracellular lactate , an increased rate of extracellular acidification , and inhibition by the non-metabolized glucose analog , 2-deoxy-D-glucose .\n\nAssociated with the induction of aerobic glycolysis was a pathway-wide induction of glycolysis gene expression , as well as protein accumulation of an established glycolysis master-regulator , hypoxia-inducible factor 1A .\n\nArsenite-induced alteration of energy production in human cells represents the type of fundamental perturbation that could extend to many tissue targets and diseases .", "output": "Cellular energetics" }, { "input": "The DNA mismatch repair ( MMR ) pathway corrects specific types of DNA replication errors that affect microsatellites and thus is critical for maintaining genomic integrity .\n\nThe genes of the MMR pathway are highly conserved across different organisms .\n\nLikewise , defective MMR function universally results in microsatellite instability ( MSI ) which is a hallmark of certain types of cancer associated with the Mendelian disorder hereditary nonpolyposis colorectal cancer .\n\n( Lynch syndrome ) .\n\nTo identify previously unrecognized deleted genes or loci that can lead to MSI , we developed a functional genomics screen utilizing a plasmid containing a microsatellite sequence that is a host spot for MSI mutations and the comprehensive homozygous diploid deletion mutant resource for Saccharomyces cerevisiae .\n\nThis pool represents a collection of non-essential homozygous yeast diploid ( 2N ) mutants in which there are deletions for over four thousand yeast open reading frames ( ORFs ) .\n\nFrom our screen , we identified a deletion mutant strain of the PAU24 gene that leads to MSI .\n\nIn a series of validation experiments , we determined that this PAU24 mutant strain had an increased MSI-specific mutation rate in comparison to the original background wildtype strain , other deletion mutants and comparable to a MMR mutant involving the MLH1 gene .\n\nLikewise , in yeast strains with a deletion of PAU24 , we identified specific de novo indel mutations that occurred within the targeted microsatellite used for this screen .", "output": "Genomic instability and mutation" }, { "input": "The current study was undertaken to study the effect of a macerated extract of Nigella sativa seeds in normal as well as in tumour bearing mice against gamma radiation-induced cellular damage to normal tissues .\n\nThis was done to mimic the clinical setting where in , normal tissues of cancer patients undergoing radiotherapy are exposed to the deleterious effects of radiation .\n\nThe protection of cellular DNA was analysed in peripheral blood leucocytes of whole body irradiated mice following pretreatment with macerated extract of Nigella sativa seeds ( 100 mg/kg ) , using alkaline comet assay , and also estimating biochemical and blood parameters such as levels of antioxidant enzymes superoxide dismutase and catalase , thiobarbituric acid reactive substances and protein oxidation in organs such as spleen , liver , brain and intestine haemoglobin and total leucocyte count , respectively .\n\nThe results showed that the macerated extract of Nigella sativa seeds protected the liver , spleen , brain and intestines both in normal as well as tumour bearing mice .\n\nThis study concludes that macerated extract of Nigella sativa seeds has protective effects against radiation-induced damage and biochemical alterations which could be attributed to the ability to scavenge free radicals and its antioxidant properties .\n\nHence macerated extract of Nigella sativa seeds , could be used in combination with radiation to protect against oxidative stress in normal tissues and improving the quality of life of cancer patients by mitigating unwanted side effects of radiation in normal tissues .", "output": "Tumor promoting inflammation, Genomic instability and mutation" }, { "input": "Previously , we reported that Helicobacter pylori-associated gastritis and gastric cancer are closely associated with increased levels of hydrogen sulfide ( H2S ) and that Korean red ginseng significantly reduced the severity of H. pylori-associated gastric diseases by attenuating H2S generation .\n\nBecause the incubation of endothelial cells with H2S has been known to enhance their angiogenic activities , we hypothesized that the amelioration of H2S-induced gastric inflammation or angiogenesis in human umbilical vascular endothelial cells ( HUVECs ) might explain the preventive effect of Korean red ginseng on H. pylori-associated carcinogenesis .\n\nThe expression of inflammatory mediators , angiogenic growth factors , and angiogenic activities in the absence or presence of Korean red ginseng extracts ( KRGE ) were evaluated in HUVECs stimulated with the H2S generator sodium hydrogen sulfide ( NaHS ) .\n\nKRGE efficiently decreased the expression of cystathionine \u03b2-synthase and cystathionine \u03b3-lyase , enzymes that are essential for H2S synthesis .\n\nConcomitantly , a significant decrease in the expression of inflammatory mediators , including cyclooxygenase-2 and inducible nitric oxide synthase , and several angiogenic factors , including interleukin ( IL)-8 , hypoxia inducible factor-1a , vascular endothelial growth factor , IL-6 , and matrix metalloproteinases , was observed ; all of these factors are normally induced after NaHS .\n\nAn in vitro angiogenesis assay demonstrated that NaHS significantly increased tube formation in endothelial cells , whereas KRGE pretreatment significantly attenuated tube formation .\n\nNaHS activated p38 and Akt , increasing the expression of angiogenic factors and the proliferation of HUVECs , whereas KRGE effectively abrogated this H2S-activated angiogenesis and the increase in inflammatory mediators in vascular endothelial cells .\n\nIn conclusion , KRGE was able to mitigate H2S-induced angiogenesis , implying that antagonistic action against H2S-induced angiogenesis may be the mechanism underlying the gastric cancer preventive effects of KRGE in H. pylori infection .", "output": "Inducing angiogenesis, Tumor promoting inflammation" }, { "input": "BACKGROUND In this paper we propose a chemical physics mechanism for the initiation of the glycolytic switch commonly known as the Warburg hypothesis , whereby glycolytic activity terminating in lactate continues even in well-oxygenated cells .\n\nWe show that this may result in cancer via mitotic failure , recasting the current conception of the Warburg effect as a metabolic dysregulation consequent to cancer , to a biophysical defect that may contribute to cancer initiation .\n\nMODEL Our model is based on analogs of thermodynamic concepts that tie non-equilibrium fluid dynamics ultimately to metabolic imbalance , disrupted microtubule dynamics , and finally , genomic instability , from which cancers can arise .\n\nSpecifically , we discuss how an analog of non-equilibrium Rayleigh-Benard convection can result in glycolytic oscillations and cause a cell to become locked into a higher-entropy state characteristic of cancer .\n\nCONCLUSIONS A quantitative model is presented that attributes the well-known Warburg effect to a biophysical mechanism driven by a convective disturbance in the cell .\n\nContrary to current understanding , this effect may precipitate cancer development , rather than follow from it , providing new insights into carcinogenesis , cancer treatment , and prevention .", "output": "Cellular energetics" }, { "input": "The green fluorescent protein ( GFP ) is the most commonly used reporter protein for monitoring gene expression and protein localization in a variety of living and fixed cells , including not only prokaryotes , but also eukaryotes , e.g. , yeasts , mammals , plants and fish .\n\nIn general , it is thought that GFP is nontoxic to cells , although there are some reports on the side effect of GFP .\n\nFurther , details of the molecular mechanism concerning the side effect of GFP remain unclear .\n\nHere we show that Ku80 , but not XRCC4 , plays an important role in the mechanism of the resistance to cytotoxicity induced by enhanced GFP ( EGFP ) .\n\nEGFP inhibited both cell proliferation and colony formation , and induced cell death in Ku80-deficient hamster cells , i.e. , xrs-6 cells .\n\nIn addition , Ku80 attenuated EGFP-induced cytotoxicity in the xrs-6 cells .\n\nNo EGFP-induced cytotoxicity was observed in the NHEJ core protein XRCC4-deficient hamster cells , i.e. , XR-1 cells .\n\nFurthermore , EGFP markedly enhanced X-ray-induced cytotoxicity in the xrs-6 cells .\n\nThese results suggest that Ku80 plays a key role in the novel NHEJ-independent defense mechanism against EGFP-induced cytotoxicity .\n\nCaution should be taken in considering of the potential influence by the stress response mechanism , namely , the Ku80-dependent elimination mechanism of EGFP-induced cytotoxicity , being activated , even when using EGFP-expressing cells in which Ku80 functions normally .", "output": "Genomic instability and mutation" }, { "input": "As a member of peroxiredoxin ( Prx ) family , PrxIII is predominantly located in mitochondria and plays an important role as a scavenger of reactive oxygen species ( ROS ) .\n\nSince previous reports demonstrated over-expression of PrxIII in cervical cancer , we conducted the present study to investigate the significance of PrxIII in cervical cancer development and/or progression .\n\nCervical cancer cells were cultured from tissues derived from cervical cancer patients .\n\nAfter successful knockdown of PrxIII expression by small interfering RNA , we evaluated ROS level , viable cell number , and apoptosis of cervical cancer cells along with the culture time .\n\nThe production of ROS was increased in cervical cancer cells as compared with normal cervical epithelia .\n\nKnockdown of PrxIII expression induced up-regulation of other Prx members including PrxI , PrxII , and PrxV .\n\nROS level was higher in down-regulated cervical cancer cells than in controls and the difference was increasing with culture time .\n\nWe also observed increased apoptosis and decreased viable cell number in down-regulated cervical cancer cells .\n\nOur results suggest that PrxIII is an indispensable ROS scavenger , which protects tumor cells against oxidative damage and subsequent apoptosis .", "output": "Tumor promoting inflammation, Resisting cell death" }, { "input": "Cellular senescence , an irreversible cell cycle arrest induced by a diversity of stimuli , has been considered as an innate tumor suppressing mechanism with implications and applications in cancer therapy .\n\nUsing a targeted proteomics approach , we show that fibroblasts induced into senescence by expression of oncogenic Ras exhibit a decrease of global acetylation on all core histones , consistent with formation of senescence-associated heterochromatic foci .\n\nWe also detected clear increases in repressive markers ( e.g. >50% elevation of H3K27me2/3 ) along with decreases in histone marks associated with increased transcriptional expression/elongation ( e.g .\n\nH3K36me2/3 ) .\n\nDespite the increases in repressive marks of chromatin , 179 loci ( of 2206 total ) were found to be upregulated by global quantitative proteomics .\n\nThe changes in the cytosolic proteome indicated an upregulation of mitochondrial proteins and downregulation of proteins involved in glycolysis .\n\nThese alterations in primary metabolism are opposite to the well-known Warburg effect observed in cancer cells .\n\nThis study significantly improves our understanding of stress-induced senescence and provides a potential application for triggering it in antiproliferative strategies that target the primary metabolism in cancer cells .", "output": "Enabling replicative immortality, Cellular energetics" }, { "input": "BACKGROUND Methyl sulfone is a small molecule that reverses cancerous phenotypes of a melanoma cell line .\n\nHere , we sought to determine whether methyl sulfone was effective against human breast cancer tissue .\n\nMETHODS We studied normal and cancerous breast tissue obtained from 17 patients .\n\nRESULTS Methyl sulfone introduced structural order , with cancer tissue taking on the morphology of normal in vivo breast tissue ; this structural order was sustainable over long-term culture .\n\nMethyl sulfone promoted proper wound healing , including migration of cells into wounded areas and establishment of stable contact inhibition once wounds were covered .\n\nMethyl sulfone decreased expression of two breast stem cell marker proteins , HCAM and OCT3/4 , which are associated with aberrantly rapid migration of metastatic cells .\n\nFinally , normal and cancerous primary breast cells remained viable and healthy in methyl sulfone culture for at least 90 days .\n\nCONCLUSION Methyl sulfone reintroduced a normal structural phenotype to human breast cancer tissues .", "output": "Evading growth suppressors, Activating invasion and metastasis" }, { "input": "BACKGROUND Matrix metalloproteinases comprise a family of enzyme degrade components of extra cellular matrix .\n\nThere are single nucleotide polymorphisms in the promoter regions of several genes with ability to influence cancer susceptibility .\n\nThe aim of this study was to analyze association between MMP3 promoter polymorphisms and colorectal cancer occurrence and progression .\n\nMATERIALS AND METHODS In this case-control study 120 colorectal cancer patients and 100 controls were genotyped using polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP ) on the genomic deoxyribonucleic acid ( DNA ) .\n\nThe patients group was divided into different subgroups : a subgroup without metastatic activity ( M(-) ) and a subgroup that had developed metastasis ( M(+) ) .\n\nRESULTS There was a significant difference in frequency of the MMP-3 genotype between cases and controls ( \\u03c7\\u0382 = 16.17 ; P = 0.0003 ) .\n\nThe 5A homozygote in patients and controls was significantly different .\n\nThe frequency of the 5A allele among affected patients ( 67.91% ) was significantly higher than among the healthy controls ( 49% ; \\u03c7(2) = 16.17 , P = 0.00005 ) .\n\nAt the time of diagnosis , individual who was carrying the 5A allele was more represented in the M(+) subgroup than in M(-) subgroup ( \u03c7\u00b2 = 7.49 ; P = 0.006 , OR = 3.86 ; 95% CI , 1.43-10.33 ) .\n\nThe difference between M(-) and controls did not observe statistically significant ( \u03c7\u00b2 = 0.009 ; P = 0.92 ) .\n\nCONCLUSIONS Our results suggest that the presence of 5A polymorphism at the MMP-3 promoter region may favor the growth and the metastasis process in colorectal cancer patients and could be looked at as a risk factor for a worse prognosis .", "output": "Activating invasion and metastasis" }, { "input": "Hypoxia-inducible factor-1 ( HIF-1 ) plays a critical role in reprogramming cancer metabolism toward aerobic glycolysis ( i.e. , the Warburg effect ) , which is critical to supplying cancer cells with the biomass needed for proliferation .\n\nPrevious studies have shown that cetuximab , an EGF receptor-blocking monoclonal antibody , downregulates the alpha subunit of HIF-1 ( HIF-1\\u03b1 ) through the inhibition of EGF receptor downstream cell signaling and that downregulation of HIF-1\\u03b1 is required for cetuximab-induced antiproliferative effects .\n\nHowever , the mechanism underlying these actions has yet to be identified .\n\nIn this study , we used the Seahorse XF96 extracellular flux analyzer to assess the effect of cetuximab treatment on changes in glycolysis and mitochondrial respiration , the two major energy-producing pathways , in live cells .\n\nWe found that cetuximab downregulated lactate dehydrogenase A ( LDH-A ) and inhibited glycolysis in cetuximab-sensitive head and neck squamous cell carcinoma ( HNSCC ) cells in an HIF-1\\u03b1 downregulation-dependent manner .\n\nHNSCC cells with acquired cetuximab resistance expressed a high level of HIF-1\\u03b1 and were highly glycolytic .\n\nOverexpression of a HIF-1\\u03b1 mutant ( HIF-1\\u03b1/\\u0394ODD ) conferred resistance to cetuximab-induced G1 phase cell-cycle arrest , which could be overcome by knockdown of LDH-A expression .\n\nInhibition of LDH-A activity with oxamate enhanced the response of cetuximab-resistant cells to cetuximab .\n\nCetuximab had no noticeable inhibitory effect on glycolysis in nontransformed cells .\n\nThese findings provide novel mechanistic insights into cetuximab-induced cell-cycle arrest from the perspective of cancer metabolism and suggest novel strategies for enhancing cetuximab response .", "output": "Sustaining proliferative signaling, Cellular energetics" }, { "input": "Oncogene-induced senescence is now recognized as being an important mechanism in protecting against cancer .\n\nThe phenotypic consequences , i.e. , the inhibition of cell proliferation , are well described in model systems and specific events/key players defined .\n\nHowever , there is still the need to understand , at a more global level , the network of events involved both in terms of cause and consequence .\n\nThis paper shows the power of systematic proteomic analyses , both targeted and global , in addressing such biological questions , highlighting the widespread nature of histone acetylation changes , and the opposite metabolic changes to those seen in the Warburg effect .", "output": "Cellular energetics" }, { "input": "CONTEXT Papillary thyroid carcinoma ( PTC ) is the most frequent thyroid tumor and is responsible for the overall increase in thyroid cancer incidence .\n\nS100A11 ( calgizzarin ) , a member of the S100 Ca(2+)-binding protein family , is involved in several different biological processes .\n\nS100A11 has been found up-regulated in PTC , both at the mRNA and protein levels .\n\nOBJECTIVE Through a combination of expression analysis and functional in vitro and in vivo studies , we have attempted to gain insight into the relevance of S100A11 overexpression in PTC biology .\n\nDESIGN The expression of the S100A11 gene in PTC was investigated in several gene expression data sets .\n\nThe effect of S100A11 silencing on the hallmarks of the malignant phenotype of several PTC-derived cell lines was investigated .\n\nIn NIH3T3 cells , the cooperation of S100A11 with the different PTC-specific oncogenes was assessed .\n\nRESULTS We found that the S100A11 gene expression is frequently up-regulated in PTC , anaplastic thyroid carcinoma , but not in follicular thyroid carcinoma .\n\nS100A11 overexpression was also detected in PTC-derived cell lines , which were then used for functional studies .\n\nS100A11 silencing in PTC-derived cell lines did not affect cell proliferation , whereas it reduced the loss of contact inhibition , anchorage-independent growth , and resistance to anoikis .\n\nCotransfection experiments in NIH3T3 cells showed that overexpression of the S100A11 gene was able to enhance the transforming capabilities of the different PTC-associated oncogenes by affecting the loss of contact inhibition , anchorage-independent growth , and in vivo tumor formation .\n\nCONCLUSION Our data indicate that S100A11 overexpression exerts a protumoral functional role in PTC pathogenesis .", "output": "Evading growth suppressors" }, { "input": "BACKGROUND Yes Associated Protein ( YAP ) has been implicated in the control of organ size by regulating cell proliferation and survival .\n\nYAP is a transcriptional coactivator that controls cellular responses through interaction with TEAD transcription factors in the nucleus , while its transcriptional functions are inhibited by phosphorylation-dependent translocation to the cytosol .\n\nYAP overexpression has been associated with different types of cancer , such as lung , skin , prostate , ovary and liver cancer .\n\nRecently , YAP was linked to E-cadherin-dependent regulation of contact inhibition in breast cancer cells .\n\nRESULTS In this study we examined YAP protein expression and cellular localization in 237 cases of human invasive breast cancer by immunohistochemistry and related its expression to clinicopathological features and E-cadherin expression .\n\nWe observed that invasive lobular carcinoma is characterized by higher expression levels of both nuclear and cytosolic YAP ( p\u2009<\u20090.001 ) .\n\nNuclear YAP expression did not associate with other variables such as lymph node involvement , tumor grade , tumor size , mitotic activity or the molecular sub-types of invasive breast cancer .\n\nWe observed that high nuclear and cytosolic YAP expression are associated with the E-cadherin deficient breast cancer subtype ILC ( p\u2009<\u20090.001 ) and cell lines derived from human breast cancers and conditional mouse models of human lobular breast cancer .\n\nCONCLUSIONS Since our data indicate that nuclear YAP localization is more common in breast cancers lacking functional adherens junctions , it suggests that YAP-mediated transcription may be involved in the development and progression of invasive lobular breast cancer .", "output": "Activating invasion and metastasis" }, { "input": "Key cellular decisions , such as proliferation or growth arrest , typically occur at spatially defined locations within tissues .\n\nLoss of this spatial control is a hallmark of many diseases , including cancer .\n\nYet , how these patterns are established is incompletely understood .\n\nHere , we report that physical and architectural features of a multicellular sheet inform cells about their proliferative capacity through mechanical regulation of YAP and TAZ , known mediators of Hippo signaling and organ growth .\n\nYAP/TAZ activity is confined to cells exposed to mechanical stresses , such as stretching , location at edges/curvatures contouring an epithelial sheet , or stiffness of the surrounding extracellular matrix .\n\nWeidentify the F-actin-capping/severing proteins Cofilin , CapZ , and Gelsolin as essential gatekeepers that limit YAP/TAZ activity in cells experiencing low mechanical stresses , including contact inhibition of proliferation .\n\nWe propose that mechanical forces are overarching regulators of YAP/TAZ in multicellular contexts , setting responsiveness to Hippo , WNT , and GPCR signaling .", "output": "Evading growth suppressors" }, { "input": "The aim of this study was to investigate the inhibitory effects of rutaecarpine on DNA strand breaks and apoptosis induced by hydrogen peroxide ( H2O2 ) in murine Hepa-1c1c7 cells .\n\nOxidative DNA damage was estimated by nuclear condensation assessment , fluorescence-activated cell sorting analysis , and Comet assay .\n\nRutaecarpine inhibited cell death induced by 500 \\u03bcM H2O2 , as assessed by 4',6-diamidino-2-phenylindole ( DAPI ) staining .\n\nTreatment with rutaecarpine reduced the number of DNA strand breaks induced by H2O2 , as assessed by DAPI staining and Comet assay , and increased quinone reductase , phosphatidylinositol 3-kinase , and pAkt protein levels , as assessed by western blotting .", "output": "Genomic instability and mutation, Resisting cell death" }, { "input": "Although the immense efforts have been made for cancer prevention , early diagnosis , and treatment , cancer morbidity and mortality has not been decreased during last forty years .\n\nEspecially , lung cancer is top-ranked in cancer-associated human death .\n\nTherefore , effective strategy is strongly required for the management of lung cancer .\n\nIn the present study , we found that novel daphnane diterpenoids , yuanhualine ( YL ) , yuanhuahine ( YH ) and yuanhuagine ( YG ) isolated from the flower of Daphne genkwa ( Thymelaeaceae ) , exhibited potent anti-proliferative activities against human lung A549 cells with the IC50 values of 7.0 , 15.2 and 24.7 nM , respectively .\n\nFlow cytometric analysis revealed that the daphnane diterpenoids induced cell-cycle arrest in the G0/G1 as well as G2/M phase in A549 cells .\n\nThe cell-cycle arrests were well correlated with the expression of checkpoint proteins including the up-regulation of cyclin-dependent kinase inhibitor p21 and p53 and down-regulation of cyclin A , cyclin B1 , cyclin E , cyclin dependent kinase 4 , cdc2 , phosphorylation of Rb and cMyc expression .\n\nIn the analysis of signal transduction molecules , the daphnane diterpenoids suppressed the activation of Akt , STAT3 and Src in human lung cancer cells .\n\nThe daphnane diterpenoids also exerted the potent anti-proliferative activity against anticancer-drug resistant cancer cells including gemcitabine-resistant A549 , gefitinib- , erlotinib-resistant H292 cells .\n\nSynergistic effects in the growth inhibition were also observed when yuanhualine was combined with gemcitabine , gefitinib or erlotinib in A549 cells .\n\nTaken together , these findings suggest that the novel daphnane diterpenoids might provide lead candidates for the development of therapeutic agents for human lung cancers .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "Epidermal growth factor receptor ( EGFR ) gene amplification and overexpression are commonly present in glioblastoma , and confer advantages of growth , invasiveness and radio/chemotherapy-resistance for tumour cells .\n\nHere , we assessed the role of EGFR activation for downstream mitogenic signalling in the commonly used glioblastoma cell line U251 .\n\nDespite the high expression level , activation of EGFR under standard culture conditions was low .\n\nIntact EGFR function was verified by the rapid phosphorylation of EGFR and downstream mitogen-activated protein(MAP) kinase ERK1/2 upon addition of exogenous EGF to serum-starved cells .\n\nBy contrast , addition of fetal bovine serum ( FBS ) activated downstream ERK1/2 via the MAP kinase kinase without phosphorylating EGFR .\n\nA phosphoreceptor tyrosine kinase array showed FBS-induced activation of insulin-like growth factor-1 receptor ( IGF-1R),and the IGF-1R inhibitor AG1024 inhibited FBS-induced phosphorylation of ERK1/2 , implying IGF-1R as the major driver of FBS-associated mitogenic signalling in the absence of exogenous EGF .\n\nThese findings have important implications for in vitro drug testing in glioblastoma .\n\nMoreover , activation of ERK1/2 was also strongly influenced by growth state and cell density of U251 cultures .\n\nRe-seeding exponentially growing cultures at high cell density induced p27/CDKN1B expression and suppressed P-ERK1/2 indicating a certain regulation of proliferation by contact inhibition .\n\nStrikingly , highly activated ERK1/2 signalling and cell cycle progression occurred when cells were released from plateau phase regardless of high seeding density .\n\nThis phenomenon might implicate a proliferation response in the early recurrence observed after clinical therapy in glioblastoma patients .\n\nHowever , whether it will recapitulate in vivo remains to be demonstrated .", "output": "Sustaining proliferative signaling, Evading growth suppressors" }, { "input": "Accumulating data suggest arsenic may be an endocrine disruptor and tentatively linked to breast cancer by some studies .\n\nTherefore , we tested the effects of chronic inorganic arsenic exposure on the normal estrogen receptor ( ER)-negative breast epithelial cell line , MCF-10A .\n\nCells were chronically exposed to a low-level arsenite ( 500nM ) for up to 24weeks .\n\nMarkers of cancer cell phenotype and the expression of critical genes relevant to breast cancer or stem cells ( SCs ) were examined .\n\nAfter 24weeks , chronic arsenic-exposed breast epithelial ( CABE ) cells showed increases in secreted MMP activity , colony formation , invasion , and proliferation rate , indicating an acquired cancer cell phenotype .\n\nThese CABE cells presented with basal-like breast cancer characteristics , including ER-\u03b1 , HER-2 , and progesterone receptor negativity , and overexpression of K5 and p63 .\n\nPutative CD44(+)/CD24(-/low) breast SCs were increased to 80% over control in CABE cells .\n\nCABE cells also formed multilayer cell mounds , indicative of loss of contact inhibition .\n\nThese mounds showed high levels of K5 and p63 , indicating the potential presence of cancer stem cells ( CSCs ) .\n\nEpithelial-to-mesenchymal transition occurred during arsenic exposure .\n\nOverexpression of aromatase , a key rate-limiting enzyme in estrogen synthesis , occurred with arsenic starting early on in exposure .\n\nLevels of 17\u03b2-estradiol increased in CABE cells and their conditioned medium .\n\nThe aromatase inhibitor letrozole abolished arsenic-induced increases in 17\u03b2-estradiol production and reversed cancer cell phenotype .\n\nThus , chronic arsenic exposure drives human breast epithelia into a cancer cell phenotype with an apparent overabundance of putative CSCs .\n\nArsenic appears to transform breast epithelia through overexpression of aromatase , thereby activating oncogenic processes independent of ER .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Activating invasion and metastasis" }, { "input": "Metabolic reprogramming in cancer is manifested by persistent aerobic glycolysis and suppression of mitochondrial function and is known as the Warburg effect .\n\nThe Warburg effect contributes to cancer progression and is considered to be a promising therapeutic target .\n\nUnderstanding the mechanisms used by cancer cells to suppress their mitochondria may lead to development of new approaches to reverse metabolic reprogramming .\n\nWe have evaluated mitochondrial function and morphology in poorly respiring LM7 and 143B osteosarcoma ( OS ) cell lines showing the Warburg effect in comparison with actively respiring Saos2 and HOS OS cells and noncancerous osteoblastic hFOB cells .\n\nIn LM7 and 143B cells , we detected markers of the mitochondrial permeability transition ( MPT ) , such as mitochondrial swelling , depolarization , and membrane permeabilization .\n\nIn addition , we detected mitochondrial swelling in human OS xenografts in mice and archival human OS specimens using electron microscopy .\n\nThe MPT inhibitor sanglifehrin A reversed MPT markers and increased respiration in LM7 and 143B cells .\n\nOur data suggest that the MPT may play a role in suppression of mitochondrial function , contributing to the Warburg effect in cancer .", "output": "Cellular energetics" }, { "input": "Cancer cells require glucose to support their rapid growth through a process known as aerobic glycolysis , or the Warburg effect .\n\nAs in ovarian cancer cells , increased metabolic activity and glucose concentration has been linked to aggressiveness of cancer .\n\nHowever , it is unclear as to whether targeting the glycolytic pathway may kill the malignant cells and likely have broad therapeutic implications against ovarian cancer metastasis .\n\nIn the present research , we found that EF24 , a HIF-1\\u03b1 inhibitor , could significantly block glucose uptake , the rate of glycolysis , and lactate production compared with vehicle treatment in SKOV-3 , A2780 and OVCAR-3 cells .\n\nThese results might possibly contribute to the further observation that EF24 could inhibit ovarian cancer cell migration and invasion from wound healing and Transwell assays .\n\nFurthermore , as an important mediator of glucose metabolism , glucose transporter 1 ( Glut1 ) was found to contribute to the function of EF24 in both energy metabolism and metastasis .\n\nTo examine the effect of EF24 and the mediated role of Glut1 in vivo in a xenograph subcutaneous tumor model , intraperitoneal metastasis and lung metastasis model were introduced .\n\nOur results indicated that EF24 treatment could inhibit tumor growth , intraperitoneal metastasis and lung metastasis of SKOV-3 cells , and Glut1 is a possible mediator for the role of EF24 .\n\nIn conclusion , our results highlight that an anti-cancer reagent with an inhibiting effect on energy metabolism could inhibit metastasis , and EF24 is a possible candidate for anti-metastasis therapeutic applications for ovarian cancer .", "output": "Cellular energetics, Activating invasion and metastasis" }, { "input": "One classical feature of cancer cells is their metabolic acquisition of a highly glycolytic phenotype .\n\nCarbon monoxide ( CO ) , one of the products of the cytoprotective molecule heme oxygenase-1 ( HO-1 ) in cancer cells , has been implicated in carcinogenesis and therapeutic resistance .\n\nHowever , the functional contributions of CO and HO-1 to these processes are poorly defined .\n\nIn human prostate cancers , we found that HO-1 was nuclear localized in malignant cells , with low enzymatic activity in moderately differentiated tumors correlating with relatively worse clinical outcomes .\n\nExposure to CO sensitized prostate cancer cells but not normal cells to chemotherapy , with growth arrest and apoptosis induced in vivo in part through mitotic catastrophe .\n\nCO targeted mitochondria activity in cancer cells as evidenced by higher oxygen consumption , free radical generation , and mitochondrial collapse .\n\nCollectively , our findings indicated that CO transiently induces an anti-Warburg effect by rapidly fueling cancer cell bioenergetics , ultimately resulting in metabolic exhaustion .", "output": "Cellular energetics, Resisting cell death" }, { "input": "Metastasis is the most lethal attribute of human malignancy .\n\nHigh-level expression of survivin is involved in both carcinogenesis and angiogenesis in cancer .\n\nPrevious studies indicate that a mutation of the threonine residue at position 34 ( Thr34Ala ) of survivin generates a dominant-negative mutant that induces apoptosis , inhibits angiogenesis , and suppresses highly metastatic breast carcinoma in mouse models .\n\nWe investigated the efficacy of gene therapy with a survivin dominant-negative mutant and possible factors related to lymph node metastasis .\n\nThe metastasis rate was compared between each group in order to find a survivin-targeted therapy against lymphangiogenesis in its earliest stages .\n\nWe established lymph node metastasis models and treated animals with H22 tumors with Lip-mSurvivinT34A ( Lip-mS ) , Lip-plasmid ( Lip-P ) , or normal saline ( NS ) .\n\nEight days after the last dose , five randomly chosen mice from each group were sacrificed .\n\nWe detected the apoptotic index , microvessel density ( MVD ) , lymphatic microvessel density ( LMVD ) , and the expression of VEGF-D with immunohistochemistry .\n\nAfter the remaining animals were sacrificed , we compared the tumor-infiltrated lymph nodes in each group .\n\nAdministration of mSurvivinT34A plasmid complexed with cationic liposome ( DOTAP/chol ) resulted in the efficacious inhibition of tumor growth and lymph node metastasis within the mouse H22 tumor model .\n\nThese responses were associated with tumor cell apoptosis , and angiogenesis and lymphangiogenesis inhibition .\n\nOur results suggested that Lip-mSurvivinT34A induced apoptosis and inhibited tumor angiogenesis and lymphangiogenesis , thus suppressing tumor growth and lymphatic metastasis .\n\nThe mSurvivinT34A survivin mutant is a promising strategy of gene therapy to inhibit lymphatic metastasis .", "output": "Inducing angiogenesis, Resisting cell death, Activating invasion and metastasis" }, { "input": "Pyruvate kinase M2 ( PKM2 ) is a key player in the Warburg effect of cancer cells .\n\nHowever , the mechanisms of regulating PKM2 are not fully elucidated .\n\nHere , we identified the protein-serine/threonine kinase PIM2 , a known oncogene , as a novel binding partner of PKM2 .\n\nThe interaction between PIM2 and PKM2 was confirmed by multiple biochemical approaches in vitro and in cultured cells .\n\nImportantly , we found that PIM2 could directly phosphorylate PKM2 on the Thr-454 residue , resulting in an increase of PKM2 protein levels .\n\nCompared with wild type , PKM2 with the phosphorylation-defective mutation displayed a reduced effect on glycolysis , co-activating HIF-1\\u03b1 and \\u03b2-catenin , and cell proliferation , while enhancing mitochondrial respiration of cancer cells .\n\nThese findings demonstrate that PIM2-dependent phosphorylation of PKM2 is critical for regulating the Warburg effect in cancer , highlighting PIM2 as a potential therapeutic target .", "output": "Genomic instability and mutation, Cellular energetics" }, { "input": "Glioblastoma multiforme is the most aggressive primary brain tumor in adults .\n\nOverexpression of the EGF receptor ( EGFR ) is recognized as a widespread oncogenic signature in glioblastoma multiforme , but the complexity of its contributions is not fully understood , nor the most effective ways to leverage anti-EGFR therapy in this setting .\n\nHypoxia is known to drive the aggressive character of glioblastoma multiforme by promoting aerobic glycolysis rather than pyruvate oxidation carried out in mitochondria ( OXPHOS ) , a phenomenon termed the Warburg effect , which is a general feature of oncogenesis .\n\nIn this study , we report that hypoxia drives expression of the pyruvate dehydrogenase kinase ( PDK1 ) and EGFR along with the hypoxia-inducing factor ( HIF)-1\\u03b1 in human glioblastoma multiforme cells .\n\nPDK1 is a HIF-1-regulated gene and our findings indicated that hypoxia-induced PDK1 expression may promote EGFR activation , initiating a feed-forward loop that can sustain malignant progression .\n\nRNAi-mediated attenuation of PDK1 and EGFR lowered PDK1-EGFR activation and decreased HIF-1\\u03b1 expression , shifting the Warburg phenotype to OXPHOS and inhibiting glioblastoma multiforme growth and proliferation .\n\nIn clinical specimens of glioblastoma multiforme , we found that immunohistochemical expression of PDK1 , EGFR , and HIF-1\\u03b1 were elevated in glioblastoma multiforme specimens when compared with normal brain tissues .\n\nCollectively , our studies establish PDK1 as a key driver and candidate therapeutic target in glioblastoma multiforme .", "output": "Sustaining proliferative signaling, Cellular energetics" }, { "input": "BACKGROUND Anti-angiogenic treatment of glioblastoma characteristically results in therapy resistance and tumor progression via diffuse infiltration .\n\nMonitoring tumor progression in these patients is thwarted because therapy results in tumor invisibility in contrast-enhanced ( CE ) MRI .\n\nTo address this problem , we examined whether tumor progression could be monitored by metabolic mapping using ( 1)H MR spectroscopic imaging ( MRSI ) .\n\nMETHODS We treated groups of BALB/c nu/nu mice carrying different orthotopic diffuse-infiltrative glioblastoma xenografts with bevacizumab ( anti-vascular endothelial growth factor [ VEGF ] antibody , n = 13 ) , cabozantinib ( combined VEGF receptor 2/c-Met tyrosine kinase inhibitor , n = 11 ) , or placebo ( n = 15 ) and compared CE-MRI with MRS-derived metabolic maps before , during , and after treatment .\n\nMetabolic maps and CE-MRIs were subsequently correlated to histology and immunohistochemistry .\n\nRESULTS In vivo imaging of choline/n-acetyl aspartate ratios via multivoxel MRS is better able to evaluate response to therapy than CE-MRI .\n\nLactate imaging revealed that diffuse infiltrative areas in glioblastoma xenografts did not present with excessive glycolysis .\n\nIn contrast , glycolysis was observed in hypoxic areas in angiogenesis-dependent compact regions of glioma only , especially after anti-angiogenic treatment .\n\nCONCLUSION Our data present MRSI as a powerful and feasible approach that is superior to CE-MRI and may provide handles for optimizing treatment of glioma .\n\nFurthermore , we show that glycolysis is more prominent in hypoxic areas than in areas of diffuse infiltrative growth .\n\nThe Warburg hypothesis of persisting glycolysis in tumors under normoxic conditions may thus not be valid for diffuse glioma .", "output": "Inducing angiogenesis, Cellular energetics" }, { "input": "BACKGROUND Alcohol consumption promotes hepatocellular carcinoma ( HCC ) .\n\nThe responsible mechanisms are not well understood .\n\nHepatocarcinogenesis increases with age and is enhanced by factors that impose a demand for liver regeneration .\n\nBecause alcohol is hepatotoxic , habitual alcohol ingestion evokes a recurrent demand for hepatic regeneration .\n\nThe alcohol-preferring ( P ) rat model mimics the level of alcohol consumption by humans who habitually abuse alcohol .\n\nPreviously , we showed that habitual heavy alcohol ingestion amplified age-related hepatocarcinogenesis in P rats , with over 80% of alcohol-consuming P rats developing HCCs after 18months of alcohol exposure , compared with only 5% of water-drinking controls .\n\nMETHODS Herein , we used quantitative real-time PCR and quantitative immunocytochemistry to compare liver tissues from alcohol-consuming P rats and water-fed P rat controls after 6 , 12 , or 18months of drinking .\n\nWe aimed to identify potential mechanisms that might underlie the differences in liver cancer formation and hypothesized that chronic alcohol ingestion would activate Hedgehog ( HH ) , a regenerative signaling pathway that is overactivated in HCC .\n\nRESULTS Chronic alcohol ingestion amplified age-related degenerative changes in hepatocytes , but did not cause appreciable liver inflammation or fibrosis even after 18months of heavy drinking .\n\nHH signaling was also enhanced by alcohol exposure , as evidenced by increased levels of mRNAs encoding HH ligands , HH-regulated transcription factors , and HH target genes .\n\nImmunocytochemistry confirmed increased alcohol-related accumulation of HH ligand-producing cells and HH-responsive target cells .\n\nHH-related regenerative responses were also induced in alcohol-exposed rats .\n\nThree of these processes ( i.e. , deregulated progenitor expansion , the reverse Warburg effect , and epithelial-to-mesenchymal transitions ) are known to promote cancer growth in other tissues .\n\nCONCLUSIONS Alcohol-related changes in Hedgehog signaling and resultant deregulation of liver cell replacement might promote hepatocarcinogenesis .", "output": "Tumor promoting inflammation, Cellular energetics, Activating invasion and metastasis" }, { "input": "Effective glucose diet : We report the development and activity of glucose-conjugated LDH-A inhibitors designed for dual targeting of the Warburg effect ( elevated glucose uptake and glycolysis ) in cancer cells .\n\nGlycoconjugation could be applied to inhibitors of many enzymes involved in glycolysis or tumor metabolism .", "output": "Cellular energetics" }, { "input": "Metastatic breast tumors undergo epithelial-to-mesenchymal transition ( EMT ) , which renders them resistant to therapies targeted to the primary cancers .\n\nThe mechanistic link between mtDNA ( mitochondrial DNA ) reduction , often seen in breast cancer patients , and EMT is unknown .\n\nWe demonstrate that reducing mtDNA content in human mammary epithelial cells ( hMECs ) activates Calcineurin ( Cn)-dependent mitochondrial retrograde signaling pathway , which induces EMT-like reprogramming to fibroblastic morphology , loss of cell polarity , contact inhibition and acquired migratory and invasive phenotype .\n\nNotably , mtDNA reduction generates breast cancer stem cells .\n\nIn addition to retrograde signaling markers , there is an induction of mesenchymal genes but loss of epithelial markers in these cells .\n\nThe changes are reversed by either restoring the mtDNA content or knockdown of CnA\u03b1 mRNA , indicating the causal role of retrograde signaling in EMT .\n\nOur results point to a new therapeutic strategy for metastatic breast cancers targeted to the mitochondrial retrograde signaling pathway for abrogating EMT and attenuating cancer stem cells , which evade conventional therapies .\n\nWe report a novel regulatory mechanism by which low mtDNA content generates EMT and cancer stem cells in hMECs.Oncogene advance online publication , 4 November 2013 ; doi:10.1038/onc.2013.467 .", "output": "Evading growth suppressors, Activating invasion and metastasis" }, { "input": "Cancers are characterized by an increasing glycolytic activity , which is called the Warburg effect .\n\nAlthough this phenomenon is well known , the mechanism of the enhanced rate of glycolysis in cancer has not yet been clearly recognized .\n\nThe present study investigated the glycolytic rate , regulatory enzymatic activities and the expression of phosphofructokinase-1 ( PFK-1 ) in human breast cancer and paracancer tissues .\n\nHuman breast cancer tissues have an increased degree of glycolytic efficiency and regulatory enzymatic activities , which have been shown in previous studies .\n\nHowever , the present study identified a number of novel observations .\n\nThe total PFK-1 levels were higher in human breast cancer tissues than in paracancer tissues , and further investigations revealed differential PFK-1 isoenzyme expression patterns between human breast cancer and paracancer tissues .\n\nThe human breast cancer and paracancer tissues mainly expressed PFK-P and PFK-L isoforms , respectively .\n\nLinear-regression analysis showed that , depending on the pathological stage of breast cancer , the expression of PFK-P was significantly positively correlated with the activity of PFK-1 .\n\nThus , during the development of human breast cancer , the enhancement of glycolytic activity depends primarily on the conversion of the PFK-1 , from PFK-L to PFK-P .", "output": "Cellular energetics" }, { "input": "In normal tissues , strict control of tissue size is achieved by regulating cell numbers .\n\nThe mechanism that controls total cell number is known as contact inhibition of growth and it depends on the NF2/Merlin pathway .\n\nNegative regulation of this pathway by deleterious mutations or by oncogenes results in cell transformation and tumor progression .\n\nHere we provide evidence that the CD43 sialomucin cooperates with oncogenic signals to promote cell transformation by abrogating the contact inhibition of growth through a molecular mechanism that involves AKT-dependent Merlin phosphorylation and degradation .\n\nAccordingly , inhibition of endogenous CD43 expression by RNA interference in lung , cervix and colon human cancer cells impaired tumor growth in vivo .\n\nThese data underscore a previously unidentified role for CD43 in non-hematopoietic tumor progression .", "output": "Evading growth suppressors" }, { "input": "BACKGROUND Aerobic glycolysis , namely the Warburg effect , is the main hallmark of cancer cells .\n\nMitochondrial respiratory dysfunction has been proposed to be one of the major causes for such glycolytic shift .\n\nThis hypothesis has been revisited as tumors appear to undergo waves of gene regulation during progression , some of which rely on functional mitochondria .\n\nIn this framework , the role of mitochondrial complex I is still debated , in particular with respect to the effect of mitochondrial DNA mutations in cancer metabolism .\n\nThe aim of this work is to provide the proof of concept that functional complex I is necessary to sustain tumor progression .\n\nMETHODS Complex I-null osteosarcoma cells were complemented with allotopically expressed complex I subunit 1 ( MT-ND1 ) .\n\nComplex I re-assembly and function recovery , also in terms of NADH consumption , were assessed .\n\nClones were tested for their ability to grow in soft agar and to generate tumor masses in nude mice .\n\nHypoxia levels were evaluated via pimonidazole staining and hypoxia-inducible factor-1\\u03b1 ( HIF-1\\u03b1 ) immunoblotting and histochemical staining. 454-pyrosequencing was implemented to obtain global transcriptomic profiling of allotopic and non-allotopic xenografts .\n\nRESULTS Complementation of a truncative mutation in the gene encoding MT-ND1 , showed that a functional enzyme was required to perform the glycolytic shift during the hypoxia response and to induce a Warburg profile in vitro and in vivo , fostering cancer progression .\n\nSuch trigger was mediated by HIF-1\\u03b1 , whose stabilization was regulated after recovery of the balance between \\u03b1-ketoglutarate and succinate due to a recuperation of NADH consumption that followed complex I rescue .\n\nCONCLUSION Respiratory complex I is essential for the induction of Warburg effect and adaptation to hypoxia of cancer cells , allowing them to sustain tumor growth .\n\nDifferently from other mitochondrial tumor suppressor genes , therefore , a complex I severe mutation such as the one here reported may confer anti-tumorigenic properties , highlighting the prognostic values of such genetic markers in cancer .", "output": "Genomic instability and mutation, Cellular energetics" }, { "input": "BACKGROUND Multiple myeloma ( MM ) is a fatal plasma cell malignancy exhibiting enhanced glucose consumption associated with an aerobic glycolytic phenotype ( i.e. , the Warburg effect ) .\n\nWe have previously demonstrated that myeloma cells exhibit constitutive plasma membrane ( PM ) localization of GLUT4 , consistent with the dependence of MM cells on this transporter for maintenance of glucose consumption rates , proliferative capacity , and viability .\n\nThe purpose of this study was to investigate the molecular basis of constitutive GLUT4 plasma membrane localization in MM cells .\n\nFINDINGS We have elucidated a novel mechanism through which myeloma cells achieve constitutive GLUT4 activation involving elevated expression of the Rab-GTPase activating protein AS160_v2 splice variant to promote the Warburg effect .\n\nAS160_v2-positive MM cell lines display constitutive Thr642 phosphorylation , known to be required for inactivation of AS160 Rab-GAP activity .\n\nImportantly , we show that enforced expression of AS160_v2 is required for GLUT4 PM translocation and activation in these select MM lines .\n\nFurthermore , we demonstrate that ectopic expression of a full-length , phospho-deficient AS160 mutant is sufficient to impair constitutive GLUT4 cell surface residence , which is characteristic of MM cells .\n\nCONCLUSIONS This is the first study to tie AS160 de-regulation to increased glucose consumption rates and the Warburg effect in cancer .\n\nFuture studies investigating connections between the insulin/IGF-1/AS160_v2/GLUT4 axis and FDG-PET positivity in myeloma patients are warranted and could provide rationale for therapeutically targeting this pathway in MM patients with advanced disease .", "output": "Cellular energetics" }, { "input": "ANXA2 , a member of the annexin family , is overexpressed and plays important roles in tumor development .\n\nHowever , the significance of ANXA2 expression in gastric carcinoma has not been clarified.To elucidate its roles in growth of gastric cancer , ANXA2 expression in SGC-7901 cells was inhibited with a designated siRNA , then cell proliferation , cell cycling , apoptosis and motility were determined by MTT assay , flow cytometry , Hoechst 33342 staining and wound healing assay , respectively .\n\nTo further assess the behavior of ANXA2 deleted SGC- 7901 cells , changes of microstructures were observed under fluorescence microscopy , laser scanning confocal microscopy and electron microscopy .\n\nWe found that inhibition of ANXA2 expression caused cell proliferation to decrease significantly with G1 arrest , motility to be reduced with changes in pseudopodia/filopodia structure and F-actin and \u03b2-tubulin expression , and apoptosis to be enhanced albeit without significance .\n\nAt the same time , ANXA2 deletion resulted in fewer pseudopodia/filopodia , non-stained areas were increased , contact inhibition among cells reappeared , and expression of F-actin and \u03b2-tubulin was decreased , with induction of polymerized disassembled forms .\n\nTaken together , these data suggest that ANXA2 overexpression is important to maintain the malignancy of cancer cells , and this member of the annexin family has potential to be considered as a target for the gene therapy of gastric carcinoma .", "output": "Sustaining proliferative signaling, Evading growth suppressors, Resisting cell death" }, { "input": "Hypoxia has long been linked to the Warburg effect , yet the underlying mechanism remains largely unclear .\n\nIt is also not known if lncRNAs are involved in the contribution of hypoxia to the Warburg effect .\n\nHere we show that lincRNA-p21 is a hypoxia-responsive lncRNA and is essential for hypoxia-enhanced glycolysis .\n\nHypoxia/HIF-1\\u03b1-induced lincRNA-p21 is able to bind HIF-1\\u03b1 and VHL and thus disrupts the VHL-HIF-1\\u03b1 interaction .\n\nThis disassociation attenuates VHL-mediated HIF-1\\u03b1 ubiquitination and causes HIF-1\\u03b1 accumulation .\n\nThese data indicate the existence of a positive feedback loop between HIF-1\\u03b1 and lincRNA-p21 that promotes glycolysis under hypoxia .\n\nThe ability of lincRNA-p21 to promote tumor growth is validated in mouse xenograft models .\n\nTogether , these findings suggest that lincRNA-p21 is an important player in the regulation of the Warburg effect and also implicate lincRNA-p21 as a valuable therapeutic target for cancer .", "output": "Cellular energetics" }, { "input": "Aims : RAS-induced tumorigenesis has been suggested to follow a three-stage model consisting of an initial RAS activation , senescence induction , and evasion of p53-dependent senescence checkpoints .\n\nWhile reactive oxygen species ( ROS ) act as second messengers in RAS-induced senescence , they are also involved in oncogenic transformation by inducing proliferation and promoting mutations .\n\nIn the current work , we investigated the role of extracellular superoxide dismutase ( SOD3 ) in RAS-induced senescence and immortalization in vitro and in vivo .\n\nWe used a mouse embryonic fibroblast ( MEF ) primary cell model together with immortalized and transformed human cell lines derived from papillary and anaplastic thyroid cancer .\n\nResults : Based on our data , sod3 RNA interference in H-RasV12-transduced cells markedly inhibited cell growth , while sod3 over-expression in MEFs initially caused a proliferative burst followed by the activation of DNA damage checkpoints , induction of p53-p21 signal transduction , and senescence .\n\nSubsequently , sod3-transduced MEF cells developed co-operative p21-p16 down-regulation and acquired transformed cell characteristics such as increased telomerase activity , loss of contact inhibition , growth in low-nutrient conditions , and in vivo tumorigenesis .\n\nInterestingly , as reported previously with RAS , we showed a dose-dependent response to SOD3 in vitro and in vivo involving transcriptional and non-transcriptional regulatory mechanisms .\n\nInnovation : SOD3 may mediate H-RasV12-induced initiation of primary cell immortalization .\n\nConclusions : Our results indicate that SOD3 influences growth signaling in primary and cancer cells downstream of the ras oncogene and could serve as a therapy target at an early tumorigenesis phase .", "output": "Genomic instability and mutation, Enabling replicative immortality, Sustaining proliferative signaling, Evading growth suppressors, Tumor promoting inflammation" }, { "input": "Tumour cells primarily utilize aerobic glycolysis for energy production , a phenomenon known as the Warburg effect .\n\nIts mechanism is not well understood .\n\nThe tumour suppressor gene p53 is frequently mutated in tumours .\n\nMany tumour-associated mutant p53 ( mutp53 ) proteins not only lose tumour suppressive function but also gain new oncogenic functions that are independent of wild-type p53 , defined as mutp53 gain of function ( GOF ) .\n\nHere we show that tumour-associated mutp53 stimulates the Warburg effect in cultured cells and mutp53 knockin mice as a new mutp53 GOF .\n\nMutp53 stimulates the Warburg effect through promoting GLUT1 translocation to the plasma membrane , which is mediated by activated RhoA and its downstream effector ROCK .\n\nInhibition of RhoA/ROCK/GLUT1 signalling largely abolishes mutp53 GOF in stimulating the Warburg effect .\n\nFurthermore , inhibition of glycolysis in tumour cells greatly compromises mutp53 GOF in promoting tumorigenesis .\n\nThus , our results reveal a new mutp53 GOF and a mechanism for controlling the Warburg effect .", "output": "Cellular energetics" }, { "input": "Our previous study demonstrated that 5-aminolevulinic acid ( ALA ) administered to mice stimulates oxidative phosphorylation by upregulation of the mitochondrial respiratory chain complex IV enzyme cytochrome c oxidase ( COX ) .\n\nThe present study investigated whether ALA disrupts the Warburg effect , which represents a shift in ATP generation from oxidative phosphorylation to glycolysis , protecting tumor cells against oxidative stress-mediated apoptosis .\n\nThe human lung carcinoma cell line A549 exposed to ALA exhibited enhanced oxidative phosphorylation , which was indicated by an increase in COX protein expression and oxygen consumption .\n\nFurthermore , ALA suppressed glycolysis-mediated acidosis .\n\nThis normalization of the ATP metabolic pathways significantly increased the generation of superoxide anion radical ( O2\\u2022- ) and the functional expression of active caspase-3 , leading to caspase-dependent apoptosis .\n\nThese data demonstrate that ALA inhibits the Warburg effect and induces cancer cell death .\n\nUse of this endogenous compound might constitute a novel approach to cancer therapy .", "output": "Cellular energetics, Resisting cell death" }, { "input": "Ceramide is a sphingolipid metabolite that induces cancer cell death .\n\nWhen C6-ceramide is encapsulated in a nanoliposome bilayer formulation , cell death is selectively induced in tumor models .\n\nHowever , the mechanism underlying this selectivity is unknown .\n\nAs most tumors exhibit a preferential switch to glycolysis , as described in the \" Warburg effect \" , we hypothesize that ceramide nanoliposomes selectively target this glycolytic pathway in cancer .\n\nWe utilize chronic lymphocytic leukemia ( CLL ) as a cancer model , which has an increased dependency on glycolysis .\n\nIn CLL cells , we demonstrate that C6-ceramide nanoliposomes , but not control nanoliposomes , induce caspase 3/7-independent necrotic cell death .\n\nNanoliposomal ceramide inhibits both the RNA and protein expression of GAPDH , an enzyme in the glycolytic pathway , which is overexpressed in CLL .\n\nTo confirm that ceramide targets GAPDH , we demonstrate that downregulation of GAPDH potentiates the decrease in ATP after ceramide treatment and exogenous pyruvate treatment as well as GAPDH overexpression partially rescues ceramide-induced necrosis .\n\nFinally , an in vivo murine model of CLL shows that nanoliposomal C6-ceramide treatment elicits tumor regression , concomitant with GAPDH downregulation .\n\nWe conclude that selective inhibition of the glycolytic pathway in CLL cells with nanoliposomal C6-ceramide could potentially be an effective therapy for leukemia by targeting the Warburg effect .", "output": "Cellular energetics, Resisting cell death" }, { "input": "High-throughput screening of a small-molecule library identified a 5-triazolo-2-arylpyridazinone as a novel inhibitor of the important glycolytic enzyme 6-phosphofructo-2-kinase/2,6-bisphosphatase 3 ( PFKFB3 ) .\n\nSuch inhibitors are of interest due to PFKFB3's control of the important glycolytic pathway used by cancer cells to generate ATP .\n\nA series of analogues was synthesized to study structure-activity relationships key to enzyme inhibition .\n\nChanges to the triazolo or pyridazinone rings were not favoured , but limited-size substitutions on the aryl ring provided modest increases in potency against the enzyme .\n\nSelected analogues and literature-described inhibitors were evaluated for their ability to suppress the glycolytic pathway , as detected by a decrease in lactate production , but none of these compounds demonstrated such suppression at non-cytotoxic concentrations .", "output": "Cellular energetics" }, { "input": "Aberrant glucose metabolism characterized by high levels of glycolysis , even in the presence of oxygen , is an important hallmark of cancer .\n\nThis metabolic reprogramming referred to as the Warburg effect is essential to the survival of tumor cells and provides them with substrates required for biomass generation .\n\nMolecular mechanisms responsible for this shift in glucose metabolism remain elusive .\n\nAs described herein , we found that aberrant expression of the proinflammatory protein transglutaminase 2 ( TG2 ) is an important regulator of the Warburg effect in mammary epithelial cells .\n\nMechanistically , TG2 regulated metabolic reprogramming by constitutively activating nuclear factor ( NF)-\\u03baB , which binds to the hypoxia-inducible factor ( HIF)-1\\u03b1 promoter and induces its expression even under normoxic conditions .\n\nTG2/NF-\\u03baB-induced increase in HIF-1\\u03b1 expression was associated with increased glucose uptake , increased lactate production and decreased oxygen consumption by mitochondria .\n\nExperimental suppression of TG2 attenuated HIF-1\\u03b1 expression and reversed downstream events in mammary epithelial cells .\n\nMoreover , downregulation of p65/RelA or HIF-1\\u03b1 expression in these cells restored normal glucose uptake , lactate production , mitochondrial respiration and glycolytic protein expression .\n\nOur results suggest that aberrant expression of TG2 is a master regulator of metabolic reprogramming and facilitates metabolic alterations in epithelial cells even under normoxic conditions .\n\nA TG2-induced shift in glucose metabolism helps breast cancer cells to survive under stressful conditions and promotes their metastatic competence .", "output": "Tumor promoting inflammation, Cellular energetics, Activating invasion and metastasis" } ] }