【摘要】 目的探讨microRNA-222(miR-222)通过调控Akt信号通路对内皮祖细胞(EPCs)迁移功能的影响及对靶向调控转录因子ETS-1(ETS proto-oncogene 1,transcription factor)的影响。方法从正常成人外周静脉血分离、体外培养后鉴定EPCs。分别向人内皮祖细胞(hEPCs)转染人工合成的miR-222的模拟物(mimic)、抑制物(inhibitor)和相应的阴性对照(NC)。用荧光定量PCR法测miR-222及ETS-1 mRNA的表达。应用Western blot测量每组的ETS-1、p-Akt和Akt的蛋白表达。各组EPCs用Transwell检测其迁移。结果miR-222可以靶向影响ETS-1的mRNA和蛋白表达。与对照组比较,上调miR-222表达可致ETS-1的mRNA和蛋白表达降低,同时抑制EPCs的迁移能力(均P<0.05)。相反,下调miR-222表达可致ETS-1的mRNA和蛋白表达升高,同时增强EPCs的迁移能力(均P<0.05)。Western blot检测显示,上调miR-222表达抑制p-Akt的蛋白水平;下调miR-222表达增强p-Akt的蛋白水平(均P<0.05)。结论miR-222可通过调控Akt信号通路和靶向转录因子ETS-1影响EPCs的迁移功能。更多还原

【Abstract】 Objective To explore the effects of microRNA-222(miR-222) on the migration of endothelial progenitor cells(EPCs) and the role of Akt signaling pathway and its target gene, ETS-1(ETS proto-oncogene 1, transcription factor), in this process. Methods EPCs were respectively isolated, cultured and identified from human peripheral blood. EPCs from healthy donors were transfected with miR-222 mimic, mimic negative control(NC), miR-222 inhibitor, or inhibitor NC.The expression levels of ETS-1 mRNA and miR-222 were measured by qRT-PCR. The protein levels of ETS-1, Akt and p-Akt were analyzed by western blot. The migratory ability of EPCs was assessed using Transwell assay. Results MiR-222 can regulate the expression of ETS-1 mRNA and protein.Over-expression of miR-222 significantly down-regulated the expression of ETS-1 in EPCs at the mRNA and protein levels and inhibited migratory ability of EPCs(all P < 0. 05). Furthermore,knockdown of miR-222 significantly up-regulated the expression of ETS-1 in EPCs at the mRNA and protein levels and promoted migratory ability of EPCs(all P < 0. 05). Western blot showed that up-regulation of miR-222 reduced the levels of p-Akt. Down-regulation of miR-222 increased the levels of p-Akt( all P< 0. 05). Conclusion MiR-222 regulated the migratory ability of EPCs via its target, EST-1, and through Akt signaling pathway.更多还原