miR-126对内皮祖细胞增殖和迁移的影响以及靶基因的检测

miR-126 promotes endothelial progenitor cell migration and targets KANK2

目的研究miR-126（microRNA-126）对大鼠骨髓源性内皮祖细胞（endothelialprogenitorcells，EPCs）增殖和迁移能力的影响，并探讨miR-126新的靶基因。方法采用电转的方法，在EPCs中转染对照寡核苷酸和miR-126的模拟物或抑制物。噻唑蓝（MTr）法检测细胞增殖，划痕和transwell实验检测细胞迁移能力的改变。microRNA靶基因预测软件TargentScan在线分析miR-126的靶基因，并进一步用Western blot检测靶基因的表达变化。结果（1）miR-126模拟物在转染后24h对EPCs的增殖有促进作用（P〈0．01），转染后48和72h，miR-126对EPCs增殖没有影响。（2）划痕和transwell实验证实miR-126模拟物可以促进EPCs的迁移（P〈0．01），miR-126抑制物抑制EPCs的迁移（P〈0．01）。（3）TargetScan在线软件预测KANK2是miR-126的靶基因。（4）Western blot检测结果显示miR-126模拟物抑制KANK2的表达，miR-126抑制物促进KANK2的表达。结论miR-126对EPCs的增殖有一过性的促进作用，miR-126可以促进EPCs的迁移能力并靶向KANK2蛋白，抑制KANK2蛋白的表达。

Objective To investigate the role of miR-126 （micro RNA-126） in rat endothelialprogenitor cells （EPCs） proliferation and migration and the starget gene of miR-126 by bioinformatics and experimental survey. Method EPCs were transfected with control oligoes and miR-126 mimics or inhibitor by electroporation. MTT was performed to evaluate the growth of EPCs subjecting to miR-126 overexpression. Cell migration analysis was done by wound healing and transwell assay. The target genes of miR-126 were predicted by TargetScan and validated by Western blot. Result （1） miR-126 mimics promoted EPCs growth at 24 h post cell transfection （P 〈 0. 01 ）. In contrast, the EPCs growth was immue from miR-126 application at 48 and 72 h. （2） Both the wound healing and transwell assay show that miR- 126 promotes EPCs migration （ P 〈 0.01 ） and miR-126 inhibitor inhibits EPCs migration （ P 〈 0. 01 ）. （3）It is predicted that KANK2 is the potential target gene of miR-126 by TargetScan online software. （4） The results of Western blot indicated that miR-126 mimics repress the expression of KANK2 compared with NC but miR-126 inhibitor enhances KANK2 expression. Conclusions miR-126 has a transient effect on the promotion of EPCc growth, miR-126 promotes EPCs migration and targets KANK2 protein.