Silencing of Claudin-11 Is Associated with Increased Invasiveness of Gastric Cancer Cells 英文参考文献.docVIP

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Silencing of Claudin-11 Is Associated with Increased Invasiveness of Gastric Cancer Cells 英文参考文献.doc

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Silencing of Claudin-11 Is Associated with Increased Invasiveness of Gastric Cancer Cells 英文参考文献

SilencingofClaudin-11IsAssociatedwithIncreased InvasivenessofGastricCancerCells RachanaAgarwal1*,YurikoMori1,YulanCheng1,ZheJin1,AlexandruV.Olaru1,JamesP.Hamilton1, StefanDavid1,FlorinM.Selaru1,JianYang1,JohnM.Abraham1,ElizabethMontgomery2,PatriceJ. Morin3,StephenJ.Meltzer1 1Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 2Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 3Laboratory of Cellular and Molecular Biology, National Institute on Aging, Baltimore,Maryland,UnitedStatesofAmerica Abstract Background:Claudinsaremembraneproteinsthatplaycriticalrolesintightjunction(TJ)formationandfunction.Members oftheclaudingenefamilyhavebeendemonstratedtobeaberrantlyregulated,andtoparticipateinthepathogenesisof various human cancers. In the present study, we report that claudin-11 (CLDN11) is silenced in gastric cancer via hypermethylationofitspromoterregion. Methodology/PrincipalFindings:LevelsofCLDN11methylationandmRNAexpressionweremeasuredinprimarygastric cancertissues,noncancerousgastricmucosae,andcelllinesofgastricoriginusingquantitativemethylation-specificPCR (qMSP)andquantitativereversetranscriptase-PCR(qRT-PCR),respectively.Analysesofpairedgastriccancersandadjacent normal gastric tissues revealed hypermethylation of the CLDN11 promoter region in gastric cancers, and this hypermethylationwassignificantlycorrelatedwithdownregulationofCLDN11expressionvs.normaltissues.TheCLDN11 promoter region was also hypermethylated in all gastric cancer cell lines tested relative to immortalized normal gastric epithelial cells. Moreover, CLDN11 mRNA expression was inversely correlated with its methylation level. Treatment of CLDN11-nonexpressing gastric cancer cells with 5-aza-29-deoxycytidine restored CLDN11 expression. Moreover, siRNA- mediatedknockdownofCLDN11expressioninnormalgastricepithelialcellsincreasedtheirmotilityandinvasiveness. C