miRNA-206对U251细胞Cx43磷酸化表达影响的研究

Effect of miRNA-206 on the Expression of Connexin 43 Phosphorylation in U251 Cells

目的:探讨微小RNA206(mi R-206)对人胶质瘤细胞U251细胞缝隙连接蛋白43磷酸化(P-Cx43)表达水平的影响。方法:本实验分为空白对照组、阴性对照组、miR-206高表达组及miR-206低表达组四组;阴性对照组、miR-206高表达组及miR-206低表达组分别转入无意义序列质粒、miR-206质粒及miR-206-inhibition质粒,荧光显微镜下观察转染效率;采用Western blotting法检测各组U251细胞P-Cx43表达水平。结果:镜下观察各组转染效率均大于80%,miR-206高表达组Cx43磷酸化表达水平明显升高,低表达组则明显降低,与空白对照组、阴性对照组比较差异有统计学意义(P<0.05)。结论:miR-206可提高U251细胞内缝隙连接蛋白43磷酸化的表达水平。

Objective:To investigate the effect of micro RNA206(miR-206) on the expression level of gap junction protein 43 phosphorylation(P-Cx43) in human glioma cell U251. Methods:This experiment was divided into blank control group, negative control group, miR-206 high expression group and miR-206 low expression group;No significant sequence plasmid, miR-206 plasmid and miR-206-inhibition plasmid were transferred into the negative control group, miR-206 high expression group and miR-206 low expression group respectively. The transfection efficiency was observed under fluorescence microscope. Western blotting method was used to detect the expression level of P-Cx43 in U251 cells. Results:The transfection efficiency of every group was more than 80%under the microscope. The expression level of P-Cx43 in miR-206 high expression group was significantly higher than that in control group and negative control group,while low expression group was significantly decreased(P<0.05). Conclusion:miR-206 can upregulate the expression level of gap junction protein 43 phosphorylation in U251 cells.