野生型着丝粒蛋白-E基因沉默对人结肠癌HCT116细胞增殖、凋亡、迁移和侵袭能力的影响

Effect of wild type centromere protein E gene silencing on proliferation,apoptosis,migration and invasion of human colon cancer HCT116 cells

目的利用RNA干扰技术下调野生型着丝粒蛋白E(Wild type centromere protein E,CENP-EWT)基因的表达,观察CENP-EWT基因沉默对结肠癌HCT116细胞增殖、凋亡、迁移和侵袭能力的影响。方法将HCT116细胞分为空白对照组、空载体转染组和CENP-EWT shRNA转染组,转染48 h后,采用巢式PCR检测细胞中CENP-EWT基因mRNA的转录水平;MTT法检测细胞的增殖活力;Hoechst法检测细胞的凋亡比例;Transwell小室试验检测细胞的迁移和侵袭能力。结果与空白对照组和空载体转染组相比,CENP-EWT shRNA转染组可有效抑制CENP-EWT基因的转录水平(P<0.05);CENP-EWT shRNA转染组细胞的增殖活力明显下降(P<0.05);细胞凋亡比例明显增加(P<0.01),细胞的迁移和侵袭能力明显增强(P<0.01)。结论 CENP-EWT基因沉默能够抑制人结肠癌HCT116细胞增殖,诱导细胞凋亡,但同时能增强细胞的迁移和侵袭能力。

Objective To down-regulate the expression of wild type centromere protein E（CENP-EWT） by RNA interference,and observe the effect of CENP-EWT gene silencing on proliferation,apoptosis,migration and invasion of human colon cancer HCT116 cells.Methods HCT116 cells were randomly divided into blank control,empty vector control and CENP-EWT shRNA transfection groups.Forty-eight hours after transfection,the CENP-EWT mRNA level in cells was determined by nested PCR,while the proliferative activity of cells by MTT method,the apoptosis rate of cells by Hoechst methd,and the migration and invasion abilities of cells by Transwell test.Results Compared with those in blank control and empty vector control groups,the transcription level of CENP-EWT mRNA and the proliferative activity of cells in CENP-EWT shRNA transfection group decreased significantly（both P〈0.05）,while the apoptosis rate as well as migration and invasion abilities of cells increased significantly（P〈0.01）.Conclusion CENP-EWT gene silencing inhibited the proliferation and induced the apoptosis,while enhanced the migration and invasion abilities of human colon cancer HCT116 cells.