论文部分内容阅读
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 Bmi-1 gene overexpression on the proliferation of human colon cancer cell line SW480 / CD133 + cells. Methods Human colon cancer SW480 / CD133 ~ + cells were infected with plasmid pMSCV-Bmi-1 (pMSCV-Bmi-1) as observation group (pMSCV-Bmi-1 / CD133 + SW480 / CD133 + cells as empty plasmid control group (pMSCV / CD133 ~ + group); blank control group without any treatment, conventional culture. Real-time fluorescence PCR and Western blotting were used to detect the level of Bmi-1 mRNA and protein expression respectively. The cell proliferation was assayed by plate clone assay. Results The levels of Bmi-1 mRNA and protein in the observation group were significantly higher than those in the blank control group and the blank control group (all P <0.01). The colony-forming rates of observation group, empty plasmid control group and blank control group were (88.45 ± 5.79)%, (52.33 ± 3.62)% and (54.66 ± 4.03)%, respectively. Control group (all P <0.01). Conclusion The Bmi-1 gene was successfully transfected into SW480 / CD133 + cells, and it can be stably and consistently expressed at mRNA and protein levels. Overexpression of Bmi-1 can significantly enhance the colony-forming ability of SW480 / CD133 ~ + cells and promote the proliferation of SW480 / CD133 ~ + cells.