Gene Silencing of Toll-Like Receptor 2 Inhibits Proliferation of Human Liver Cancer Cells and Secretion of Inflammatory Cytokines 英文参考文献.docVIP

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Gene Silencing of Toll-Like Receptor 2 Inhibits Proliferation of Human Liver Cancer Cells and Secretion of Inflammatory Cytokines 英文参考文献.doc

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Gene Silencing of Toll-Like Receptor 2 Inhibits Proliferation of Human Liver Cancer Cells and Secretion of Inflammatory Cytokines 英文参考文献

GeneSilencingofToll-LikeReceptor2Inhibits ProliferationofHumanLiverCancerCellsandSecretion ofInflammatoryCytokines YuzhengHuang1.,BingCai2.,MingXu1,ZhiqinQiu3,YonghuiTao1,YingZhang1,JieWang1, YongliangXu1,YonghuaZhou1,JingYang1,XiaofengHan3*,QiGao1* 1Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Institute of Parasitic Diseases, Wuxi, Jiangsu, China, 2Wuxi People’sHospitalofNanjingMedicalUniversity,Wuxi,Jiangsu,China,3WuxiSecondlyPeople’sHospitalofNanjingMedicalUniversity,Wuxi,Jiangsu,China Abstract Background:Toll-likereceptors(TLRs)arekeyfactorsintheinnateimmunesystemandinitiatetheinflammatoryresponse to foreign pathogens such as bacteria, fungi and viruses. In the microenvironment of tumorigenesis, TLRs can promote inflammationandcellsurvival.Toll-likereceptor2/6(TLR2/6)signalingintumorcellsisregardedasoneofthemechanisms ofchronicinflammationbutitcanalsomediatetumorcellimmuneescapeandtumorprogression.However,theexpression ofTLR2anditsbiologicalfunctioninthedevelopmentandprogressionofhepatocarcinomahavenotbeeninvestigated. ThisstudyaimedtodeterminetheexpressionofTLRs1–10intheestablishedhumanhepatocellularcarcinomacelllineBLE- 7402,toinvestigatethebiologicaleffectofTLR2oncellgrowthandsurvival. Methods: TLR expression in BLE-7402 cells was assayed by RT-PCR, real-time PCR and flow cytometry (FCM). To further investigatethefunctionofTLR2inhepatocarcinomagrowth,BLE-7402cellsweretransfectedwithrecombinantplasmids expressingoneofthreeformsofTLR2siRNA(sh-TLR2RNAi(A,BandC)).TLR2knockdownwasconfirmedusingRT-PCR,real- timePCRandfluorescencemicroscopy.TumorcellproliferationwasmonitoredbyMTTassayandsecretedcytokinesinthe supernatantoftransfectedcellsweremeasuredbybead-basedFCM,thefunctionofTLR2siRNAwasalsoinvestigatedin vivo. Results:TheBLE-7402celllineexpressedTLRs2to10atbothmRNAandproteinlevels.TLR2wasthemosthighlyexpressed TLR.WhileallthethreesiRNAsinhibitedTLR2mRNAandproteinexpression,sh-TLR2RNAi(B)hadthestrongestk