Bcl-2短发夹状RNA对BEL-7402、Caco2细胞的生长抑制作用

目的:研究Bcl-2短发夹状RNA(short hairpin RNA,shRNA)序列的表达载体对肝癌细胞系BEL-7402和结肠癌细胞系Caco2生长的抑制作用.方法:将Bcl-2 shRNA序列克隆到携带绿色荧光蛋白基因的质粒Pgenesil-1载体,采用脂质体介导的转染方法将构建的Bcl-2shRNA表达质粒转入BEL-7402、Caco2细胞,同时设立阴性shRNA组、空载体组、脂质体组、空白组作为对照.通过倒置荧光显微镜和流式细胞仪观察细胞的转染效率,Western印迹法检测Bcl-2蛋白表达水平,MTT法测定细胞增殖情况.结果:Bcl-2 shRNA转染组、阴性shRNA组、空载体组的转染效率无显著差异.转染Bcl-2 shRNA后BEL-7402、Caco2细胞Bcl-2蛋白表达水平与转染阴性shRNA、空载体组、脂质体组和空白组相比均显著降低(P＜0.05),且Bcl-2 shRNA抑制Caco2细胞Bcl-2蛋白表达比BEL-7402细胞更为明显(P＜0.05).MTT测定显示转染Bcl-2 shRNA载体入BEL-7402、Caco2细胞在72、96 h细胞生长明显受到抑制,分别与转染阴性shRNA、空载体组、脂质体组和空白组比较,差异有显著性(P＜0.05).结论:Bcl-2 shRNA可特异性地抑制BEL-7402、Caco2细胞生长.

Inhibitory effect of Bcl-2 short hairpin RNA on growth of BEL-7402 and Caco2 cells

Objective:To investigate the effects of Bcl-2 shRNA on the growth of liver carcinoma cell line BEL-7402 and colorectal carcinoma cell line Caco2.Methods: Bcl-2 shRNA was cloned into Pgenesil-1 plasmid labeled with fluorescent protein and the product was transfected into BEL-7402 and Caco2 cells with Lipofectamine 2000.The study also included shRNA as negative control,Pgenesil-1,Lipofectamine and blank control groups.Transfected cells were visualized by inverted fluorescent microscope and then assayed by flow cytometry.The expression levels of Bcl-2 protein were assayed by Western-blot and cell proliferation was measured by MTT method.Results: There was no difference in transfection rate among cells in Bcl-2 shRNA,shRNA and Pgenesil-1 vector groups.Western-blot assay showed that the expression levels of Bcl-2 protein in BEL-7402 and Caco2 cells were significantly decreased in Bcl-2 shRNA group compared with those in other 4 groups(P<0.05).Furthermore,Bcl-2 shRNA had a stronger inhibitory effect on Caco2 cells than on BEL-7402 cells(P<0.05).MTT assay showed that Bcl-2 shRNA significantly inhibited the growth of BEL-7402 and Caco2 cells(at 72 and 96 h after treatment,respectively) compared with the other 4 groups(P<0.05).Conclusion: Bcl-2 shRNA can specifically inhibit the growth of BEL-7402 and Caco2 cells.