Bmi-1基因过表达对人正常胃黏膜上皮细胞株GES-1增殖的影响

Effect of Bmi-1 gene overexpression on proliferation of human gastric mucosal epithelial cell line GES-1

目的:探讨原癌基因B淋巴瘤莫洛尼鼠白血病病毒插入区1(Bmi-1)过表达对人正常胃黏膜上皮细胞株GES-1增殖的影响。方法:采用逆转录病毒介导转染方法将携带原癌基因Bmi-1的质粒或空质粒稳定转染GES-1细胞,通过real-time PCR及Western blotting在mRNA及蛋白水平鉴定转染效果。流式细胞术检测过表达Bmi-1对GES-1细胞周期的影响。应用CCK-8(Cell Counting Kit-8)试剂盒检测稳定转染Bmi-1对GES-1细胞增殖的影响。结果:Real-time PCR及Western blotting结果均表明成功建立稳定转染Bmi-1基因的GES-1细胞株。流式细胞术结果表明,过表达Bmi-1基因使GES-1细胞G0/G1期减少,G2/M期和S期细胞增多。生长曲线显示,过表达Bmi-1基因使GES-1细胞增殖速度明显提高。结论:过表达Bmi-1基因能调控GES-1细胞的细胞周期,促进GES-1细胞的增殖。

AIM： To investigate the effect of B lymphoma Moloney murine leukemia virus insertion region 1 （Bmi - 1 ） gene overexpression on the proliferation of a human normal gastric epithelial cell line GES - 1. METHODS ： The plasmid containing Bmi - 1 gene or empty plasmid was transfected into GES - 1 cells by retroviral mediation. The ex- pression of Bmi- 1 at mRNA and protein levels was detected by quantitative real -time PCR （qRT- PCR） analysis and Western blotting, respectively. The effect of Bmi - 1 gene overexpression on the cell cycle of GES - 1 cells was evaluated by flow cytometry. The proliferation of the stably transfected ceils was measured by Cell Counting Kit - 8. RESULTS ： The results of qRT - PCR analysis and Western blotting demonstrated that stably transfected cell line was successfully estab- lished. The results of flow cytometry analysis showed that overexpression of Bmi - 1 reduced the G0/G1 phase, arrested the cells in G2/M phase and S phase. The growth curve showed that overexpression of Bmi - I resulted in increased growth speed. CONCLUSION： Increase in Bmi - 1 gene expression regulates the cell cycle and promotes the proliferation of GES - 1 cells.