目的探究microRNA-30b(miR-30b)对大鼠血管平滑肌细胞(VSMC)迁移能力的影响。方法体外分离培养大鼠VSMC,将VSMC随机分为正常对照组(未进行转染)、miR-对照组(转染miR-NC,浓度为50 nmol/L)、miR-30b mimics组(转染miR-30b mimics,浓度为50 nmol/L)。采用实时荧光定量PCR实验检测miR-30b及钙黏附蛋白E-Cadherin的表达情况;采用划痕愈合实验检测VSMC的迁移率;采用Transwell实验检测VSMC的迁移能力;采用Western blot技术检测E-Cadherin蛋白的表达。结果①miR-30b mimics组与正常对照组和miR-对照组比较,miR-30b表达水平升高(P<0.05)。②划痕愈合实验结果显示,与正常对照组和miR-对照组相比,miR-30b mimics组VSMC的划痕修复率明显降低(P<0.05)。③Transwell实验可见,miR-30b mimics组VSMC的迁移能力较正常对照组和miR-对照组明显减弱(P<0.05)。④miR-30b mimics组E-Cadherin mRNA和蛋白的表达较正常对照组和miR-对照组明显增强(P<0.05)。结论 miR-30b上调E-Cadherin的表达,从而抑制大鼠VSMC的迁移。
Aim To investigate the effect of microRNA-30b(miR-30b)on the migration of rat vascular smooth muscle cells (VSMC). Methods Rat VSMC were isolated and cultured in vitro. VSMC were randomly divided into normal control group (no transfection), miR-NC group (transfected miR-NC concentration 50 nmol/L), miR-30b mimics group (transfected miR-30b mimics, concentration 50 nmol/L). Real-time quantitative PCR was used to detect the expression of miR-30b and E-cadherin. The mobility of VSMC was detected by scratch healing test. The migration ability of VSMC was detected by Transwell assay. The expression of E-cadherin protein was detected by Western blot. Results Compared with the normal control group and the miR-NC group, the expression level of miR-30b was increased in the miR-30b mimics group (P<0.05). The results of the scratch-healing experiment showed that the migration ability of the miR-30b mimics group was significantly weaker than that of the normal control group and the miR-NC group (P<0.05). Transwell experiments showed that the migration ability of miR-30b mimics group was weakened (P<0.05). The expression of E-cadherin mRNA and protein in miR-30b mimics group was significantly higher than that in normal control group and miR-NC group (P<0.05). Conclusion miR-30b can up-regulate the expression of E-cadherin, thereby inhibiting the migration of rat VSMC.
Chinese Journal of Arteriosclerosis