顺铂对膀胱癌细胞自噬和凋亡的作用及其相关性 .pdf

顺铂对膀胱癌细胞自噬和凋亡的作用及其相关性

刘言;谷文;印胡滨;苟欣;何卫阳

【摘要】ObjectiveToidentifywhethercisplatincaninduceautophagyof

bladdercancerT24cellsandthepossiblemechanism,andtoobservethe

relationshipbetweenoutophagyandapoptosis.MethodsMTTassaywas

appliedtoinvestigatetheeffectsofvariousconcentrationofcisplatin(0,

10,20and40μg/mL)onT24survival.TEMwasperformedtodetectthe

autophagosomeformation.Westernblotassaywasusedtoanalyzethe

expressionchangesofLC3-Ⅱ,P62andextracellularsignal-regulated

kinase(ERK1/2)andp-ERKattheproteinlevel.Theeffectsofautophagy

onthesurvivalandapoptosisofbladdercancercellswereinvestiga-

ted.ResultsDDPobservablyinhibitedproliferationofbladdercancercells

inadose-dependentmanner(P0.05),the50%inhibiting

concentration(IC50)was(30.3±2.4)μg/mL;DDPinducedautophagyof

bladdernbsp;cancercells,observablyincreasedautophagosomeinduced

byDDP;up-regulatedexpressionlevelsofLC3-Ⅱproteins(P0.05),

down-regulatedexpressionofP62proteins(P0.05);DDPincreasedthe

proteinlevelofp-ERK(P0.05);TheinhibitorofERKpathwayU0126

inhibitedDDP-inducedautophagy,asevidencedbydecreaseinthe

expressionofLC3-Ⅱproteins(P0.05).Afterinhibitionofautophagyby

WTMinDDP-treatedcells,cellviabilitywasobviouslydecreasedand

apoptosiswasincreased(P0.05);DDPcombinedwithWTMobservably

enhancedcleavageofpolyADP-ribosepolymerase1(PARP-1)and

cleaved-caspase-3whichisapop-tosisrelatedproteins

(P0.05).ConclusionsAutophagycanprotectT24cellsagainstciplatin-

inducedapop-tosis,thepossiblemechanismofautophagyistheERK

signalingpathwayisactivated.%目的：研究顺铂（DDP）能否激活人膀胱癌

T24细胞自噬以及其中可能的机制，并且探索自噬和凋亡之间相关性。方法用

MTT法检测不同浓度的顺铂（0、10、20和40μg／mL）对T24细胞的增殖作用。

透射电镜观察细胞内自噬小体；用Westernblot技术检测细胞内LC3-Ⅱ、P62

蛋白和细胞外信号调节蛋白激酶1／2（ERK1／2）和磷酸化ERK1／2蛋白的表

达量。研究自噬对于DDP处理后T24细胞增殖和凋亡的影响。结果DDP可