Bmi-1基因的过表达对人结肠癌细胞株SW480/CD133^+细胞增殖的影响  

Effect of overexpression of Bmi-1 gene on proliferation of human colon cancer cell line SW480/CD133^+

作　　者：毛海波[1] 朱国栋[1] 刘国龙[1] 关明媚[1] 李宝秀[1] 翁成荫[1] 曹小飞[1] 方喜生[1] 

机构地区：[1]广州医科大学附属广州市第一人民医院肿瘤科,广东广州510180

出　　处：《临床和实验医学杂志》2017年第1期23-26,共4页Journal of Clinical and Experimental Medicine

基　　金：广州市医药卫生科技项目(编号:20121A010001)

摘　　要：目的探讨Bmi-1基因过表达对人结肠癌细胞株SW480/CD133^+细胞增殖的影响。方法用携带Bmi-1基因的质粒(pMSCV-Bmi-1)感染人结肠癌SW480/CD133^+细胞,作为观察组(pMSCV-Bmi-1/CD133^+组);以空质粒(pMSCV)感染SW480/CD133+细胞,作为空质粒对照组(pMSCV/CD133^+组);空白对照组不作任何处理,常规培养。用实时荧光PCR和免疫印迹法分别检测细胞中Bmi-1 mRNA水平和蛋白表达的水平;平板克隆实验检测细胞增殖能力的变化。结果观察组Bmi-1 mRNA和蛋白表达的水平均较空质粒对照组和空白对照组显著增高(均P<0.01)。观察组、空质粒对照组和空白对照组的细胞克隆形成率分别为(88.45±5.79)%,(52.33±3.62)%和(54.66±4.03)%,观察组分别高于空质粒对照组和空白对照组(均P<0.01)。结论成功地将Bmi-1基因导入SW480/CD133^+细胞,并能使其在mRNA水平和蛋白水平稳定、持续地表达。Bmi-1过表达可以显著增强人结直肠癌SW480/CD133^+细胞的克隆形成能力,促进SW480/CD133^+细胞增殖。Objective To investigate the effect of overexpression of Bmi-1 gene on the proliferation of human colon cancer SW480/CD133~＋ cells. Methods The observation group（ p MSCV- Bmi-1/CD133＋group） was the SW480/CD133~＋cells which were infected with plasmid containing Bmi-1 gene（ pMSCV- Bmi- 1）. The empty plasmid contol group（ pMSCV/CD133~＋group） was the SW480/CD133~＋cells which were infected with empty plasmid（ pMSCV）. The blank control group was conventional cultured without any treatment. Real- time PCR and Western blotting were used to detect the levels of mRNA Bmi-1 and protein expression in cells. The changes of proliferation ability of cells were detected by plate clone experiment. Results The levels of mRNA Bmi-1 and protein expression in the observation group were significantly higher than those in the empty plasmid control group and the blank control group（ bothP〈0.01）. The clone formation rate of the observation group,the empty plasmid control group and the blank control group was（88.45±5.79） %,（52.33±3.62） % and（54.66±4.03） % respectively. It was higher in the observation group than that in the empty plasmid control group and the blank control group respectively（P〈0.01）.Conclusion Bmi-1 gene was successfully transfected into SW480/CD133~＋cells,and continuously stable expressing in the levels of mRNA and protein. Overexpression of Bmi-1 can significantly enhance the clonality of SW480/CD133~＋cells,and promote them proliferation.

关 键 词：结肠癌 B淋巴瘤莫洛尼鼠白血病病毒插入区-1基因 SW480细胞 增殖 

分 类 号：R735.35[医药卫生—肿瘤]