顺铂对膀胱癌细胞自噬和凋亡的作用及其相关性

发布时间：2018-03-18 19:44

本文选题：膀胱癌　切入点：顺铂　出处：《重庆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:研究顺铂(cisplatin,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:采用Western blot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(extracellular signal-regulated kinase,ERK1/2)和磷酸化ERK1/2蛋白的表达量。检测自噬抑制剂渥曼青霉素(wortmannin,WTM)抑制自噬后对膀胱癌T24凋亡的影响。结果:顺铂可显著抑制T24细胞的增殖(P0.05),并呈浓度依赖性,半数抑制浓度(IC50)为(30.3±2.4)μg/mL。DDP诱导膀胱癌T24细胞自噬活化,使LC3-II蛋白的表达明显升高(P0.05),P62蛋白的表达明显下降(P0.05);细胞自噬体数目明显增加。DDP能上调ERK1/2的磷酸化(P0.05);ERK1/2通路抑制剂U0126抑制DDP诱导的膀胱癌T24细胞自噬活化,使LC3-II蛋白的表达明显下降(P0.05)。自噬抑制剂渥曼青霉素(wortmannin,WTM)抑制自噬后明显加强DDP对T24细胞的增殖抑制和凋亡促进作用(P0.05);在DDP和WTM联合处理组,凋亡指标多聚ADP-核糖聚合酶1(PARP 1)裂解明显增强,cleaved-Caspase3明显增多(P0.05)。结论:抑制自噬活化可以增强膀胱癌T24细胞凋亡,证明顺铂诱导的自噬对膀胱癌T24细胞是保护性的,并且进一步发现了自噬活化的机制可能与ERK信号通路的激活有关。
[Abstract]:Aim: to investigate whether cisplatin DDP) can activate autophagy of human bladder cancer cell line T24 and its possible mechanism. And to explore the correlation between autophagy and apoptosis. Methods: the expression of LC3-IIP62 protein and extracellular signal-regulated kinaseERK1 / 2) and phosphorylated ERK1/2 protein were detected by Western blot technique. The expression of intracellular LC3-IIP62 protein and extracellular signal-regulated kinase1 / 2 protein and phosphorylated ERK1/2 protein were detected by Western blot technique. Results: cisplatin significantly inhibited the proliferation of T24 cells in a concentration-dependent manner. IC50 was 30.3 卤2.4 渭 g / mL DDP induced autophagy in T24 cells of bladder cancer. The expression of LC3-II protein increased significantly and the expression of P0.05 / P62 protein decreased significantly. The number of autophagy increased significantly. DDP upregulated the phosphorylation of ERK1/2 phosphorylated P0.05ERK1 / 2 pathway inhibitor U0126 inhibited DDP induced autophagy activation of bladder cancer T24 cells. The expression of LC3-II protein was significantly decreased (P 0.05). The inhibition of autophagy inhibitor, Warmensin / wortmanninnine (WTM), significantly enhanced the proliferation inhibition and apoptosis promotion of DDP on T24 cells. Apoptosis index polyADP- ribosomal polymerase 1 (PARP1) cleavage increased significantly (P 0.05). Conclusion: inhibition of autophagy activation can enhance apoptosis of T24 cells, which proves that cisplatin induced autophagy is protective to bladder cancer T24 cells. Furthermore, the mechanism of autophagy activation may be related to the activation of ERK signaling pathway.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.14

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