Akt抑制剂鱼藤素对膀胱癌细胞生长的影响

Effects of Akt inhibitor deguelin on the growth of bladder cancer cells

目的 研究Akt抑制剂鱼藤素对膀胱癌T-24细胞株的生长抑制作用,并初步探讨其可能的作用机制.方法 用MTT法检测鱼藤素对T-24细胞生长的抑制率;流式细胞术检测细胞周期的改变;DNA片段化检测鱼藤素对T-24细胞凋亡的影响;RT-PCR检测鱼藤素对MDM2和GSK3β mRNA表达的影响;Western blotting检测MDM2、GSK3β蛋白表达水平的变化.结果 鱼藤素显著抑制T-24细胞的增殖,使细胞周期主要阻滞在G0/G1期,并可诱导T-24细胞发生凋亡.RT-PCR结果显示,鱼藤素处理T-24细胞后MDM2 mRNA的表达水平下降,而GSK3 β mRNA的表达水平增加;Western blotting检测发现鱼藤素下调MDM2蛋白的表达,而上调GSK3β蛋白的表达.结论 Akt抑制剂鱼藤素能抑制T-24细胞的增殖并诱导其凋亡,其机制可能与影响Akt通路下游信号分子MDM2和GSK3β的表达相关.

Objectives To study the effects of Akt inhibitor deguelin on the growth of human bladder cancer cell lines T-24,and to elucidate the possible mechanism of such effects.Methods MTT method was used to determine growth-inhibitory effect of deguelin on human bladder cancer T-24 cells.The cell cycle was detected by flow cytometry （FCM）.Apoptosis induced by deguelin in T-24 cells was determined using DNA fragmentation assay.The expression of MDM2 and GSK3β in mRNA and protein levels were detected by RT-PCR and Western blotting respectively.Results The deguelin singnificantly inhibited cell proliferation,blocked the cell cycle in the G0/G1 phase and induced apoptosis in T-24 cells.The expression levels of MDM2 mRNA and protein were down-regulated,while the expression levels of GSK3 β mRNA and proteins were up-regulated when T-24 cell were treated by deguelin.Conclusions Deguelin can inhibit the proliferation and induce the apoptosis in T-24 human bladder cancer cells.These effects may be related with its function in down-regulating MDM2 and up-regulating GSK3 β,both of them are down-stream signal meleculars in the AKt pathway.