We aimed to investigate the mechanism and effects of autophagy on cisplatin (DDP)-induced apoptosis in human gastric cancer cell line SGC7901. After SGC7901 cells were treated with DDP and/or chloroquine, cell proliferation was measured using MTT assay; cell apoptosis was determined by flow cytometry; autophagy and apotosis-related proteins expression were detected by Western blot; and quantitative analysis of autophagy after monodansylcadaverine (MDC) staining was performed using fluorescence microscopy. We found after treatment with 5 mg/L DDP for 24 h, the rates of cell apoptosis were ( )%. Autophagy, characterized by an increase in the number of autophagic vesicles and the level of LC3-II protein was observed in cells treated with DDP. After inhibition of autophagy by chloroquine, the rates of cell apoptosis were increased to ( )%, and the level of Caspase-3 and P53 protein were increased, and Bcl-2 protein was decreased. Therefore, autophagy protects human gastric cancer cell line SGC7901 against DDP-induced apoptosis, inhibition of autophagy can promote apoptosis, and combination therapy with DDP and chloroquine may be a promising therapeutic strategy for gastric cancer.
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机译:我们旨在探讨自噬对顺铂(DDP)诱导人胃癌细胞系SGC7901中细胞凋亡的机制和影响。在用DDP和/或氯喹处理SGC7901细胞后,使用MTT测定法测量细胞增殖;通过流式细胞术确定细胞凋亡;蛋白质印迹检测自噬和血管缺失相关的蛋白质表达;使用荧光显微镜进行单倍碱(MDC)染色后自噬的定量分析。我们发现用5mg / L DDP处理24小时后,细胞凋亡率为()%。自噬,其特征在于用DDP处理的细胞中观察到自噬囊泡的数量和LC3-II蛋白的水平。氯喹抑制自噬后,细胞凋亡的速率增加到()%,并且增加了Caspase-3和P53蛋白的水平,并且Bcl-2蛋白减少。因此,自噬保护人胃癌细胞系SGC7901针对DDP诱导的细胞凋亡,抑制自噬能促进凋亡,与DDP和氯喹的联合治疗可能是胃癌的有希望的治疗策略。
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