miR-222靶向BBC-3基因对肝癌细胞增殖凋亡的调控研究

Regulation of miR-222 Targeting BBC-3 Gene on Proliferation and Apoptosis of Hepatocellular Carcinoma Cells

目的探讨miR-222靶向BBC-3基因对肝癌细胞增殖凋亡的调控。方法用miR-222转染肝癌细胞HepG,分为miR-222转染组、miR-222 inhibitor转染组及空载体转染组,采用RT-PCR法检测转染后各组细胞miR-222和BBC-3 mRNA的表达水平;MTT法检测各组细胞的增殖水平以及流式细胞仪检测细胞周期及细胞凋亡率。结果RT-PCR结果表明miR-222转染组与空载体和miR-222 inhibitor转染组相比miR-222基因mRNA的表达水平显著下降,差异有统计学意义(P<0.01);miR-222转染组与空载体和miR-222 inhibitor转染组相比肝癌细胞HepG生长明显受到抑制;流式细胞仪检测结果表明空载体转染组、miR-222 inhibitor转染组、miR-222转染组三组细胞的凋亡率分别为2.65%、2.72%和13.58%,miR-222转染组与空载体和miR-222 inhibitor转染组相比细胞凋亡率显著增加,差异有统计学意义(P<0.01)。结论肝癌细胞转染miR-222基因后不但可靶向上调BBC-3基因的表达,而且降低肝癌细胞的增殖能力,从而促进细胞的凋亡。

Objective To investigate the regulation of miR-222 targeting BBC-3 gene on proliferation and apoptosis of hepatoma cells.Methods HepG cells were transfected with miR-222 and divided into 2 groups:miR-222 transfection group,miR-222 inhibitor transfection group and empty vector transfection group.The miR-genes were detected by RT-PCR.miR-222 and BBC-3 mRNA expression levels;MTT assay cell proliferation in each group and flow cytometry to detect cell cycle and apoptosis rate.Results RT-PCR results showed that miR-222 mRNA expression was significantly decreased in the miR-222 transfection group compared with the empty vector and miR-222 inhibitor transfection group(P<0.01);Compared with the empty vector and miR-222 inhibitor transfection group,the growth of HepG cells in miR-222 transfection group was significantly inhibited;Flow cytometry results showed that the apoptotic rates of cells of empty vector transfection group,miR-222 inhibitor transfection group,miR-222 transfer group were 2.65%,2.72%and 13.58%,respectively,and the apoptotic rate of the miR-222 transfection group was significantly higher than that of the empty vector and miR-222 inhibitor transfection group.The difference was statistically significant.(P<0.01).Conclusion The transfection of miR-222 gene into hepatoma cells not only up-regulates the expression of BBC-3 gene,but also reduces the proliferation of hepatoma cells and promotes cell apoptosis.