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目的:研究干扰素/维甲酸诱导死亡基因(retinoid-interferon-induced mortality,GRIM-19)对结肠癌SW480细胞凋亡的影响。方法:构建GRIM-19真核表达载体(pCMV-Flag-GRIM-19),转染入SW480细胞中,Western blotting检测GRIM-19及凋亡相关蛋白Bal-xl的表达,采用Annexin-V/PI和线粒体膜电位JC-1染色检测SW480细胞的凋亡。结果:成功构建pCMV-Flag-GRIM-19真核表达载体。pCMV-Flag-GRIM-19质粒转染SW480细胞后,GRIM-19的表达上调,凋亡抑制蛋白Bcl-xl的表达则下调。转染空质粒pCMV-Flag组SW480细胞凋亡率为(7.7±1.39)%,转染pCMV-Flag-GRIM-19质粒后SW480细胞凋亡率为(15.0±2.52)%(P<0.05)。线粒体膜电位检测显示转染空质粒pCMV-Flag组SW480细胞膜电位降低细胞为(7.5±2.09)%,而转染pCMV-Flag-GRIM-19后细胞线粒体膜电位降低细胞为(17.5±3.07)%(P<0.05)。结论:GRIM-19体外转染可有效诱导结肠癌SW480细胞凋亡。
Objective: To investigate the effects of interferon-induced mortality (GRIM-19) on the apoptosis of colon cancer SW480 cells. METHODS: The GRIM-19 eukaryotic expression vector (pCMV-Flag-GRIM-19) was constructed and transfected into SW480 cells. The expression of GRIM-19 and its associated protein Bal-xl was detected by Western blotting. Annexin-V / PI And mitochondrial membrane potential JC-1 staining SW480 cells apoptosis. Results: The eukaryotic expression vector pCMV-Flag-GRIM-19 was successfully constructed. After SW480 cells were transfected with pCMV-Flag-GRIM-19 plasmid, the expression of GRIM-19 was up-regulated and the expression of Bcl-xl was down-regulated. The apoptotic rate of SW480 cells transfected with plasmid pCMV-Flag was (7.7 ± 1.39)%. The apoptosis rate of SW480 cells transfected with pCMV-Flag-GRIM-19 plasmid was (15.0 ± 2.52)% (P <0.05). Mitochondrial membrane potential assay showed that cells transfected with pCMV-Flag group had a decrease in membrane potential of SW480 cells (7.5 ± 2.09)%, while cells transfected with pCMV-Flag-GRIM-19 cells decreased (17.5 ± 3.07)%, (P <0.05). Conclusion: GRIM-19 transfection in vitro can effectively induce the apoptosis of colon cancer SW480 cells.