五行汤经钙离子/溶酶体途径介导的SGC-7901细胞凋亡

Calcium/lysosome pathway in the apoptosis of SGC-7901 cells induced by Wuxing soup

目的探索五行汤经钙离子/溶酶体途径介导的SGC-7901细胞凋亡机制。方法吖啶橙染色检测溶酶体膜完整性,胞浆蛋白免疫印迹法检测凋亡途径蛋白变化,Annexin V结合实验和CCK-8法检测不同抑制剂对细胞凋亡和存活的影响。结果凋亡细胞的溶酶体膜渗漏,溶酶体释放蛋白酶Cathepsin D和Cathepsin B至胞浆。天冬氨酸蛋白酶Cathepsin D抑制剂对五行汤诱导的细胞凋亡无影响;半胱氨酸蛋白酶Cathepsin B、L、S抑制剂可明显减少细胞凋亡(P<0.05),且对细胞的保护作用呈浓度依赖性。结论五行汤经钙离子/溶酶体途径介导SGC-7901细胞凋亡。

Objective To explore the mechanism of calcium/lysosome pathway in the apoptosis of SGC-7901 cells induced by Wuxing soup.Methods The integrity of lysosome membrane was detected by acridine orange（AO） staining,the key proteins in apoptotic pathway were tested by Western blot,and the effect of inhibitors on cell apoptotic rate and survival rate was analyzed by Annexin V-binding ELISA and CCK-8,respectively.Results The leakage of lysosome membrane was observed by AO staining.The lysosome associated apoptotic protein Cathepsin D and Cathepsin B were released into cytosol.The inhibitors of Cathepsin B,L and S exerted protection in the survival of SGC-7901 under Wuxing soup treatment in a dose-dependent manner.However,the inhibitor of Cathepsin D had no effects on apoptosis.Conclusion Wuxing soup could induce SGC-7901 cell apoptosis through calcium mediated and lysosome involved caspase-independent pathway.