亚硒酸钠通过线粒体途径诱导人胃癌SGC-7901细胞凋亡的机制  

Sodium selenite induces apoptosis of human gastric cancer SGC-7901 cells through mitochondrial-dependent pathway

作　　者：王艳[1,2] 赵上[1] 苏衍萍[1] 刘立伟[1] 王慧[1] 曲鹏[1] 

机构地区：[1]泰山医学院组织学胚胎学教研室,山东泰安271000 [2]潍坊护理职业学院解剖学教研室,山东青州262500

出　　处：《解剖学报》2016年第3期353-358,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(81572868);山东省自然科学基金(ZR2011CM042;ZR2014CM023)

摘　　要：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 study the mechanism of gastric cancer cells apoptosis induced by sodium selenite,by investigating the expression of Bax / Bcl-2,the change of mitochondrion membrane potential and cytochrome C（ Cyt C） in the process apoptosis ofof human gastric cancer SGC-7901 cells. Methods Human gastric cancer SGC-7901 cell line was added different concentrations（ 2. 5,5. 0,10. 0μmol / L） of sodium seleite for 24 hours,48hours. The expression of Bax /Bcl-2 protein was detected by immunocytochemical and Western blotting methods. SGC-7901 cells were stained by Rhodamine123 to detect changes of mitochondrion membrane potential by flow cytometry. The changes of cytoplasm Cyt C and mitochondria Cyt C protein content were detected by Western blotting. Results Immunocytochemistry and Western blotting showed that sodium selenite increased the Bax protein levels（ P〈0. 05） and reduced the Bcl-2 protein levels（ P〈0. 05） in dose-dependent manner. Flow cytometry technique results showed that sodium selenite reduced mitochondrial membrane potential（ P〈0. 05）. The results indicated that sodium selenite may induce SGC-7901 cells apoptosis by altering Bax / Bcl-2 and reducing mitochondrial membrane potential. Western blotting showed that sodium selenite increased the cytoplasm Cyt C protein levels（ P〈0. 05） and reduced the mitochondria Cyt C protein levels（ P〈0. 05）,therefore promoted the release of Cyt C to cytoplasm. Conclusion Sodium selenite can induce apoptosis of human gastric cancer SGC-7901 cells by increasing Bax and discreasing Bcl-2 gene expression,reduce the mitochondrial membrane potential,topromote Cyt C release from mitochondria to the cytoplasm,and induce human gastric cancer SGC-7901 cell apoptosis through mitochondrial-dependent pathway.

关 键 词：亚硒酸钠 胃癌 线粒体 细胞色素C BAX/BCL-2 免疫印迹法 流式细胞术 

分 类 号：R364.5[医药卫生—病理学]