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目的通过检测亚硒酸钠诱导人胃癌SGC-7901细胞系凋亡过程中Bax/Bcl-2、线粒体膜电位和细胞色素C(Cyt C)表达的变化,探讨亚硒酸钠诱导胃癌细胞凋亡的作用机制。方法将人胃癌细胞系SGC-7901细胞加入不同浓度亚硒酸钠(2.5、5.0、10.0mol/L)的培养液中培养24h、48h,免疫细胞化学法和免疫印迹法(Western blotting)检测Bax/Bcl-2蛋白的表达;以罗丹明123(rhodamine123)为细胞染色剂,采用流式细胞技术检测细胞的线粒体膜电位的变化;Western blotting检测细胞质Cyt C和线粒体Cyt C蛋白含量的变化。结果免疫细胞化学法和Western blotting检测结果显示,亚硒酸钠能够能够提高Bax蛋白的表达(P<0.05)、降低Bcl-2蛋白的表达(P<0.05),且呈剂量依懒性;流式细胞术结果显示,亚硒酸钠能够降低细胞线粒体膜电位;提示:亚硒酸钠可以通过改变Bax、Bcl-2蛋白的含量,降低线粒体的膜电位来诱导细胞凋亡。Western blotting检测结果显示,亚硒酸钠能够提高细胞质Cyt C蛋白的表达(P<0.05)并降低线粒体内Cyt C蛋白的表达(P<0.05),促使线粒体内的Cyt C向细胞质内释放。结论亚硒酸钠通过上调Bax并下调Bcl-2蛋白表达,降低线粒体膜电位,促进Cyt C从线粒体释放到细胞质,通过线粒体途径诱导人胃癌SGC-7901细胞凋亡。
Objective To investigate the changes of Bax / Bcl-2, mitochondrial membrane potential and cytochrome C expression induced by sodium selenite in human gastric cancer cell line SGC-7901 and to investigate the effect of sodium selenite on gastric cancer cell apoptosis The mechanism of action. Methods The human gastric cancer cell line SGC-7901 cells were cultured in different concentrations of sodium selenite (2.5, 5.0, 10.0 mol / L) for 24 h and 48 h. Immunocytochemistry and Western blotting were used to detect Bax / Bcl-2 protein was detected by flow cytometry. The changes of mitochondrial membrane potential were detected by flow cytometry with rhodamine123 as the staining reagent. The changes of Cyt C and mitochondrial Cyt C were detected by Western blotting. Results The results of immunocytochemistry and Western blotting showed that sodium selenite could increase the expression of Bax protein (P <0.05) and decrease the expression of Bcl-2 protein (P <0.05) Cytometry results showed that sodium selenite can reduce mitochondrial membrane potential; Tip: Sodium selenite can change the Bax, Bcl-2 protein content, reduce the mitochondrial membrane potential to induce apoptosis. Western blotting results showed that sodium selenite could increase the expression of Cyt C (P <0.05) and decrease the expression of Cyt C in the mitochondria (P <0.05), and lead to the release of Cyt C in the mitochondria to the cytoplasm. Conclusion Sodium selenite can induce the release of Cyt C from the mitochondria to the cytoplasm by up-regulating Bax and down-regulating the expression of Bcl-2, and induce the apoptosis of SGC-7901 cells through the mitochondria pathway.