三七总皂苷对人胃癌细胞株MKN-28增殖和凋亡的影响

Effects of TSPN on the Proliferation and Apoptosis of Gastric Cancer Cell Line MKN-28

目的观察三七总皂苷(Total Saponins of Panax notoginseng,TSPN)对人胃癌细胞株MKN-28增殖和凋亡的影响,探讨其诱导细胞凋亡是否涉及上调死亡受体5(DR5)的表达。方法体外培养人胃癌细胞株MKN-28,平皿克隆形成法测定细胞锚定依赖性生长能力,PI染色流式细胞术(Flow cytometry,FCM)分析细胞周期和凋亡率,Hoechst 33258染色荧光显微镜观察凋亡细胞的形态,Western Bloting检测分析药物对DR5蛋白表达的影响。结果平皿集落形成法检测显示TSPN对MKN-28细胞生长有抑制作用,且呈浓度依赖性。PI染色FCM结果表明TSPN以时间和浓度依赖性方式诱导MKN-28细胞凋亡,且使细胞周期阻滞于G1期。Hoechst 33258染色荧光显微镜观察可见核染色质凝集,凋亡细胞呈致密浓染,与对照组相比,TSPN处理后,凋亡细胞比例增加,且呈时间和浓度依赖性。Western Bloting检测显示TSPN诱导细胞凋亡的机制涉及上调死亡受体5的表达活性。结论 TSPN在体外对人胃癌MKN-28细胞具有抑制增殖和诱导凋亡的作用,初步推断TSPN诱发胃癌细胞凋亡与其上调死亡受体5的表达活性有关。

Objective To observe the effects of total Saponins of Panax notoginseng （TSPN） on proliferation and apoptosis of human gastric cancer MKN-28 cell, and to explore whether the mechanism of apoptosis involves the up-regulation of expression of death receptors-5 （DIK5）. Method Human gastric cancer MKN-28 cells were cultured in vitro, and their fixed-dependent growth ability were assayed by agar colony formation. The cell cycle and apoptosis rate were detected by PI staining and flow cytometry （FCM）. The morphology of apoptotic cells was observed by Hoechst 33258 fluorescence staining under fluorescence microscopy. The effect of drugs on the D1K5 protein expression was analyzed by Western Bloting. Result Agar colony formation assay showed that MKN-28 cell growth was inhibited by TSPN in a dose-dependent manner. PI staining FCM results showed that the TSPN induced apoptosis of MKN-28 cells in a time and concentration-dependent manner and the cell cycle was retarded at G1 phase. Nuclear chromatin condensation can be observed by fluorescence microscopy after Hoechst 33258 staining, and apop- totic cells were dense stained. Compared with control group, the proportion ofapoptotic cells increased after TSPN treatment in a time and concentration-dependent manner. Western Bloting analysis showed that apoptosis induced by TSPN involved the upregulation of death receptor-5 expression activity. Conclusion TSPN have a role in inhibiting proliferation and inducing apoptosis on. human gastric cancer MKN-28 cells in vitro. It is inferred that this induce of apoptosis may be concerned with its role in up- regulating the expression of the death receptor-5.