顺铂诱导卵巢癌SKOV3细胞自噬和凋亡的研究

Research on autophagy and apoptosis of ovarian cancer cell line SKOV3 induced by cisplatin

目的:研究顺铂诱导卵巢癌SKOV3细胞自噬和凋亡的发生情况,并探讨可能的发生机制。方法:顺铂干预卵巢癌SKOV3细胞后,用MDC染色荧光显微镜观察自噬囊泡及流式细胞仪检测自噬率;AnnexinV-FITC/PI双染色流式细胞仪检测凋亡率及TUNEL法检测凋亡指数;Western blot检测自噬蛋白Beclin1,LC3-Ⅱ和凋亡蛋白caspase9,caspase3的表达。结果:顺铂干预SKOV3细胞后,MDC阳性细胞数明显增多;自噬率提高(P<0.05);凋亡率提高(P<0.05);凋亡指数提高(P<0.05);自噬蛋白Beclin1,LC3-Ⅱ和凋亡蛋白caspase9,caspase3表达均增强(P<0.05)。结论:顺铂不仅可诱导卵巢癌SK-OV3细胞凋亡,而且可诱导卵巢癌SKOV3细胞自噬性死亡。

Objective：To investigate autophagy and apoptosis and their mechanism in ovarian cancer cell line SKOV3 induced by cisplatin.Methods：Ovarian cancer cell line SKOV3 was intervened with cisplatin,the autophagy was analyzed by fluorescent microscope and flow cytometry（FCM）with monodansylcadaverin（MDC）staining.Apoptosis rate was measured by FCM with AnnexinV-FITC/PI staining,and apoptosis index was tested by TUNEL assays.And expression of pro-Beclin1,pro-LC3-Ⅱ,pro-caspase3 and pro-caspase9 was measured by Western blot.Results：After intervened SKOV3 cells with cisplatin,cells positive stained with MDC increased and autophagic rate was higher than that of control group（P0.05）.Apoptosis rate and apoptosis index in cisplatin group were higher than those in control group（P0.05）.Expression of pro-Beclin1,pro-LC3-Ⅱ,pro-caspase3 and pro-caspase9 was increased in SKOV3 cell lines after intervened by cisplatin（P0.05）.Conclusion：Cisplatin may not only augment autophagy but also induce apoptosis in ovarian cancer cells.