v-atpase抑制剂对人肝癌hepg2细胞行为的影响与其机制

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1、遵义医学院颐上学位论文v-ATPase抑制剂对人肝痛HepG2细胞的行为的影响及其机制EfIfectofV-ATPaseinhibitiononthecellbehaviorofhumanhepatocellularcarcinomacellsHepG2anditsmechanismsABSTRACTObjective：Inapreviousstudy,wefoundthattherewasahighervacuolarATPase(V-ATPase)expressioninhumanhepatocellularcarcinoma(HCC)tissuesandtheinhibitio

2、nofV-ATPasemarkedlyretardedthegrowthofHCCinnudemicemodel，indicatingthatV-ATPasemayplayanimportantroleinthedevelopmentandprogressofHCC．Inthisstudy,wemadeaninvestigationontheeffectofV-ATPaseinhibitionontheproliferationandapoptosisofHCCcellsandunderlyingmechanisms，whichaimedtoprovidenewevidencefor

3、pHadjustmentmechanismoftumorcellasatargetforanticancerstrategiesandtoprovideexperimentalevidencefortheclinicalapplicationofV-ATPaseinhibitor,PI一230Methods：Thestudywasperformedinhumanhepatocellularcarcinoma(HCC)cellsHepG2cellsandhumannormallivercelllines．Thecellsweresynchronized．Theproliferation

4、ofcellswasevaluatedwithafineMTTassay,cellcolonyformationexperiments，andcellviabilityanalyzercellcounting．CellularDNAcyclewasexaminedwithflowcytometryAnnexinV-FITC／PIinthecellswasanalyzedbyusingflowcytometrytodetectearlyapoptosisrateandthelateapoptosisratewasexaminedbyusingTunelmethod．Theexpress

5、ionofcaspase一3proteinwasanalyzedbywesternblot．Mitochondrialmembranepotentialwasmeasuredbyusingflowcytometry．Resialts：(1)Comparedwiththecontrolgrogp，80pMPI一230，aV-ATPaseinhibitor,incubatingfor24hours，hadnosignificanteffectonthesurvivalofHepG2andL02cellsp>0．05)，but1609M，320pM，and640pMofPI一230conc

6、entration—dependentlydecreasedthesurvivalofcellsp<0．05)．(2)Comparedwithcontrols，10pMPI一230，incubatingfor14days，significantlyreducedthenumberandrateofcolonyformationinHepG2cells(尸

7、numberandrateofcolonyformationinL02cells(p>0．05)．However,PI一230(40pM一120pM)concentrationdepend—entlyreducedthenumberandrateofcolonyformationinL02cells．(3)ThenumbersofHepG2andL02cellsbycellcountingweresignificantlydecreasedafterPI一