MicroRNA-154对大鼠肝星状细胞HSC-T6中β-catenin蛋白表达的影响

目的:探讨microRNA-154(miR-154)对大鼠肝星状细胞(hepatic stellate cellc,HSC)中β-catenin蛋白表达的影响。方法:合成miR-154模拟物(miR-154 mimics)和miR-154抑制物(miR-154 inhibitor)转染HSC-T6,以其阴性对照物(mimics NC、inhibitor NC)转染细胞作为阴性对照组,不作处理的正常细胞为空白对照组(Blank);通过实时定量RT-PCR检测转染后各组细胞内的miR-154和β-catenin mRNA的相对表达,免疫印迹法(Western blotting)检测转染后各组细胞内β-catenin蛋白的表达。结果:β-catenin蛋白表达量和β-catenin mRNA表达量在转染miR-154 mimics组显著上调,与空白对照组和阴性对照组相比差异具有统计学意义,P<0.05;β-catenin蛋白表达量和β-catenin mRNA表达量在转染miR-154 inhibitor组显著下降,与空白对照组和阴性对照组相比差异具有统计学意义,P<0.05。结论:细胞内过表达miR-154能够诱导细胞中β-catenin mRNA及蛋白的过表达;抑制细胞内miR-154的表达则能够抑制细胞中β-catenin mRNA及蛋白的表达。

Effects of microRNA-154 on β-catenin expression in hepatic stellate cell HSC-T6

OBJECTIVE To investigate effects of microRNA-154 on β-catenin expression in hepatic stellate cells. METHODS MiR-154 mimics, miR-154 inhibitor and miR-154 control were transfected into HSC. No treatment normal cells were used in blank group. Expression levels of microRNA-154 and β-catenin mRNA were measured by real time quantitative PCR. Expression level of β-catenin protein was measured by Western blotting. RESULTS Expression levels of miR-154, β-catenin mRNA and β-catenin protein were increased significantly in miR-154 mimics group than in blank group and negative control group(P<0.05). Expression levels of β-catenin mRNA and β-catenin protein were declined significantly in miR-154 inhibitor group than in blank group and negative control group(P<0.05). CONCLUSION Over expression of miR-154 in cells can induce over expression of β-catenin mRNA and β-catenin protein. Inhibition of expression of miR-154 in cells can inhibit expression of β-catenin mRNA and β-catenin protein which can inhibit occurrence of liver fibrosis.