重楼总皂苷对人胃癌MKN-45细胞凋亡及Fas/FasL信号通路的影响

目的:研究重楼总皂苷(Paridis Rhizoma total saponins,PRTS)诱导人胃癌MKN-45细胞凋亡的机制.方法:在前期研究基础上,将PRTS设定3个不同质量浓度(5,10,20 mg·L-1)处理人胃癌细胞12h,采用荧光染色法观察MKN-45细胞凋亡形态学变化;流式细胞术(Annexin V/PI双染色法)测定细胞凋亡率;ELISA测定MKN-45细胞caspase-3,caspase-8酶活性;Western blotting法检测Fas,FasL蛋白表达水平.结果:在荧光显微镜下,经PRTS处理的MKN-45细胞,可见典型的凋亡形态学特征;PRTS可明显增加细胞凋亡率,且PRTS 20 mg· L-1与对照组相比凋亡率有显著差异(P＜0.01);PRTS作用于MKN-45细胞12h,与对照组比较,其caspase-3,caspase-8活性明显增高(P＜0.01),Fas,FasL蛋白表达明显增高(P＜0.01).结论:PRTS能显著诱导人胃癌细胞MKN-45凋亡,其机制可能与增加caspase-3,caspase-8活性,上调Fas和FasL的表达有关.

Effect of Paridis Rhizoma total saponins on apoptosis of human gastric cancer cell MKN-45 and Fas/FasL signaling pathway

Objective: The study aimed to test if Paridis Rhizoma total saponins (PRTS) could induce apoptosis of human gastric cancer cell MKN-45. Method: Based on the previous researches, PRTS was set by different concentrations to treat human gastric cancer cell for 12 h (5, 10, 20 mg·L^-1). Fluorescent staining methods were adopted to observe apoptotic morphological changes of MKN-45. The apoptosis rates were analyzed by flow cytometry with Annexin V-FITC/PI staining. The enzymatic activities of caspase-3 and caspase-8 were measured by ELISA. The protein levels of Fas and FasL were detected by Western blotting. Result: Under a fluorescence microscope, MKN-45 treated by PRTS was seen typical apoptotic morphological features. PRTS significantly increased the rate of apoptosis. Compared with the control group, there exsited significant differences in apoptosis rate of PRTS concentration of 20 mg·L^-1 (P<0.01);besides, the enzymatic activities of caspase-3 and caspase-8 were promoted obviously after the effect of PRTS on MKN-45 cells for 12 h(P<0.01). The protein levels of Fas and FasL in the MKN-45 were upgraded significantly. Conclusion: PRTS can induce apoptosis of human gastric cancer cell MKN-45, which is concerned with caspase-3 and caspase-8 and upgraded Fas and FasL.