环孢素A诱导人胃癌MGC80-3细胞凋亡以及对细胞色素C蛋白表达的影响  

Effect of cyclosporin A on apoptosis and protein expression of cytochrome C in human gastric adenocarcinoma MGC80-3 cells

作　　者：黄军祥[1] 胡咏梅 石海[3] 徐雄飞[1] 周春立[1] Huang Junxiang;Hu Yongmei;Shi Hai(Dept of Gastroenterology,North Area of Suzhou Municipal Hospital,Suzhou 215008;Dept of Research,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Gastroenterology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]苏州市立医院北区消化科,苏州215008 [2]安徽医科大学第一附属医院科研处,合肥230022 [3]安徽医科大学第一附属医院消化内科,合肥230022

出　　处：《安徽医科大学学报》2019年第5期727-729,734,共4页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金青年科学基金项目(编号:11040606Q24)

摘　　要：目的研究环孢素A(CsA)诱导人胃癌MGC80-3细胞凋亡以及对细胞色素C(cytochrome C)蛋白表达的影响,探讨细胞凋亡与cytochrome C蛋白表达的关系。方法利用Annexin V-FITC/PI双染法流式细胞仪(FCM)检测不同浓度CsA处理MGC80-3细胞48 h后细胞凋亡率,透射电子显微镜观察细胞凋亡的形态学改变以及线粒体的损伤,Western blot法检测细胞内cytochrome C蛋白的表达。结果在0～20μmol/L浓度范围内CsA可显著诱导MGC80-3细胞凋亡,细胞凋亡率随CsA浓度依赖性增加,与对照组比较差异有统计学意义(F=18.64,P<0.05,P<0.01)。电镜下可见与对照组相比细胞发生核染色质固缩等典型的凋亡特征改变以及线粒体损伤。Western blot表明CsA浓度依赖性上调cytochrome C蛋白的表达(P<0.05,P<0.01)。结论 CsA能诱导MGC80-3细胞凋亡,其机制可能与线粒体损伤和上调细胞内cytochrome C蛋白表达有关。Objective To study the effect of cyclosporine A(CsA) on apoptosis of human gastric adenocarcinoma MGC80-3 cells and the protein expression of cytochrome C,and further explore the relationship between them. Methods Annexin V-FITC/PI double staining was employed to detect cell apoptosis rate by flow cytometry (FCM) after different concentration of CsA treating with MGC80-3 cells for 48 h,and utilizing transmission electron microscope to observe the morphological changes and mitochondrial damage;Western blot was applied to examine protein expression of cytochrome C. Results FCM analysis showed that cell apoptosis rate was increased with the concentration of CsA in a dose-dependent manner from the range of 0~20 μmol/L and had significant difference compared with the control group ( F =18.64, P <0.05, P <0.01). Under electron microscope,typical apoptotic changes such as nuclear chromatin condensation and mitochondrial damage were observed in the cells compared with the control group. Western blot also showed that the expression of cytochrome C protein was up-regulated in a dose-dependent manner ( P <0.05, P <0.01). Conclusion CsA may induce the apoptosis in MGC80-3 cells,and its mechanism of apoptosis may be related to up-regulation of cytochrome C protein expression and mitochondrial damage in cells.

关 键 词：环孢素A MGC80-3细胞 细胞色素C 细胞凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]