RNA干扰MAD2基因对细胞增殖的影响  

Effect of RNA interference targeting MAD2 gene on cell proliferation

作　　者：施琼[1,2] 王箭[1] 舒朝忠[1] 翁亚光[1,3] 王应雄[4] 徐远久[1] 蒋洪彦[1] 刘子杰[1] 刘青松[1] 蔡燕[1] 

机构地区：[1]重庆医科大学临床检验诊断学省部共建教育部实验室 [2]重庆医科大学基础医学院核医学教研室,重庆400016 [3]重庆医科大学感染性疾病分子生物学省部共建教育部重点实验室,重庆400016 [4]重庆医科大学公共卫生学院

出　　处：《解剖学杂志》2007年第4期400-404,共5页Chinese Journal of Anatomy

基　　金：国家自然科学基金项目(30371485);重庆市科委重点项目(渝科发计字[2004-47])

摘　　要：目的:探讨MAD2对HepG2细胞周期的影响。方法:构建针对MAD2基因的shRNA表达载体,MAD2基因与GFP融合的绿色荧光蛋白真核表达质粒pmad2-EGFP共转染HepG2细胞株,用Western印迹法检测shRNA的抑制效应;用MTT法测定细胞增殖率;用流式细胞仪检测细胞周期。结果:shRNA的重组质粒表达载体能明显抑制MAD2绿色荧光融合蛋白的表达。HepG2细胞转染有效干扰质粒48h后,细胞增殖抑制率达65%。有效干扰质粒引起G0/G1期和S期细胞的明显降低,G2/M期细胞增多。结论:在细胞水平,可通过RNA干扰特异性抑制MAD2基因来抑制细胞的增殖,为研究MAD2基因在细胞增殖中的作用机制奠定基础。Objective： To investigate the effect of MAD2 gene on cell proliferation by RNA interference. Methods： Recombinant shRNA plasmids were constructed targeting MAD2 gene. The recombinant shRNA plasmids and MAD2- GFP fusion plasmid were co-transfected into HepG2 cells, and then the expression of GFP in transfected cells was observed. Inhibitory effect of shRNAs was demonstrated by Western blot; cell proliferation was detected by MTT assay; and cell cycle was assessed by FCM. Results： The recombinant shRNA plasmids significantly inhibited MAD2-GFP fusion protein expression, and inhibited HepG2 cells proliferation to 65% at 48 h after transfection. Efficient shRNA decreased Go/G1 and S phase cells, and increased G2/M phase cells. Conclusion： At the cell level, cells proliferation can be inhibited by RNA interference to MAD2 gene expression, providing the foundation to study the function of MAD2 in cell proliferation.

关 键 词：MAD2基因 RNA干扰 细胞增殖 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]