慢病毒介导的FOXJ1过表达对胃腺癌细胞凋亡及线粒体膜电位的影响

Effects of lentivirus mediated FOXJ1 over-expression on cell apoptosis and mitochondrial membrane potential in gastric adenocarcinoma cells

目的:探讨慢病毒介导的叉头框转录因子J1(FOXJ1)过表达对胃腺癌细胞凋亡及线粒体膜电位的影响。方法:人胃腺癌细胞SGC-7901感染FOXJ1过表达慢病毒(Lv-FOXJ1),Real-time PCR和Western blot法检测细胞中FOXJ1 mRNA和蛋白的表达,MTT法检测细胞存活率,细胞克隆实验检测克隆形成率,流式细胞术检测细胞凋亡率,Western blot法检测细胞中激活型Caspase-3(C-Caspase-3)、激活型Caspase-9(C-Caspase-9)和胞浆中细胞色素C(Cyt C)蛋白表达水平,JC-1法检测线粒体膜电位。以感染阴性对照慢病毒和未感染细胞为阴性对照和空白对照。结果:FOXJ1组细胞中FOXJ1 mRNA和蛋白表达水平均增高,细胞存活率、克隆形成率降低,细胞凋亡率升高,细胞中C-Caspase-3、C-Caspase-9蛋白表达水平升高,线粒体膜电位下降,胞浆中Cyt C蛋白表达水平升高(P <0. 05)。结论:FOXJ1能够通过线粒体途径诱导胃腺癌细胞凋亡。

Aim: To investigate the effects of lentivirus mediated FOXJ1 over-expression on cell apoptosis and mitochondrial membrane potential in gastric adenocarcinoma cells. Methods: Human gastric adenocarcinoma cell SGC-7901 was infected with FOXJ1 over-expression lentivirus( Lv-FOXJ1),Real-time PCR and Western blot were used to detect the expressions of FOXJ1 mRNA and protein,cell viability was detected by MTT method,cell clone assay was used to detect clone formation ability,cell apoptosis was detected by flow cytometry,Western blot was used to detect the levels of CCaspase-3 and C-Caspase-9 protein in the cells and Cyt C in the cytoplasm,JC-1 method was used to detect the mitochondrial membrane potential. The cells infected with negative control lentivirus or without infection were the negative control or blank control. Results: Compared with those of the negative control group and blank control group,the expression of FOXJ1 in FOXJ1 group increased,the cell viability and the clone formation ability decreased,the apoptosis rate increased,the levels of C-Caspase-3 and C-Caspase-9 proteins in cells increased,mitochondrial membrane potential dropped,and the level of Cyt C protein in the cytoplasm increased( P < 0. 05). Conclusion: FOXJ1 can induce apoptosis of gastric adenocarcinoma cells through the mitochondrial pathway.