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目的观察顺铂对Hela细胞内质网应激和凋亡的影响及其机制。方法体外培养宫颈癌Hela细胞,通过MTT法测得顺铂最适应激剂量为4.5μmol/L,实验分为对照组、顺铂4 h组、顺铂8 h组、顺铂12 h组,以观察顺铂作用Hela细胞后,其内质网应激及相关凋亡的动态变化。Western Blot法检测各组细胞内质网应激、凋亡相关蛋白表达。流式细胞术检测凋亡。结果与对照组相比,内质网应激相关蛋白GRP78,内质网应激信号凋亡调节蛋白IRE1α、p-JNK表达水平明显升高,自噬相关蛋白LC3-Ⅱ表达增加;内质网应激诱导的凋亡蛋白Caspase-4和凋亡执行分子Caspase-3的裂解形式蛋白表达水平明显升高。流式细胞术检测结果显示,顺铂作用后与对照组相比,细胞凋亡率明显增加。结论顺铂可能诱导Hela细胞发生内质网应激,从而促进细胞凋亡的发生。
Objective To observe the effect of cisplatin on endoplasmic reticulum stress and apoptosis in Hela cells and its mechanism. Methods Hela cells were cultured in vitro. MTT assay showed that the optimum dose of cisplatin was 4.5 μmol / L. The experiment was divided into 4 groups: cisplatin 4 h, cisplatin 8 h and cisplatin 12 h To observe the dynamic changes of endoplasmic reticulum stress and related apoptosis in cisplatin-treated Hela cells. Western Blot method was used to detect the endoplasmic reticulum stress and the expression of apoptosis related proteins in each group. Flow cytometry was used to detect apoptosis. Results Compared with the control group, the endoplasmic reticulum stress-related protein GRP78, the expression of endoplasmic reticulum stress signaling regulatory proteins IRE1α and p-JNK were significantly increased and the expression of autophagy-related protein LC3-Ⅱ was increased. The endoplasmic reticulum Stress-induced apoptosis protein Caspase-4 and apoptosis executive molecule Caspase-3 cleavage form of protein expression increased significantly. Flow cytometry results showed that compared with the control group, cisplatin significantly increased the rate of apoptosis. Conclusion Cisplatin may induce endoplasmic reticulum stress in Hela cells and promote apoptosis.