CAJ | 학술논문

目的研究miR-126(micro RNA-126)对大鼠骨髓源性内皮祖细胞(endothelial progenitor cells,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(MTT)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24 h对EPCs的增殖有促进作用(P＜0.01),转染后48和72 h,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.
목적 연구miR-126(micro RNA-126)대대서골수원성내피조세포(endothelial progenitor cells,EPCs)증식화천이능력적영향,병탐토miR-126신적파기인.방법 채용전전적방법,재EPCs중전염대조과핵감산화miR-126적모의물혹억제물.새서람(MTT)법검측세포증식,화흔화transwell실험검측세포천이능력적개변.microRNA파기인예측연건TargentScan재선분석miR-126적파기인,병진일보용Western blot검측파기인적표체변화.결과 (1) miR-126모의물재전염후24 h대EPCs적증식유촉진작용(P＜0.01),전염후48화72 h,miR-126대EPCs증식몰유영향.(2)화흔화transwell실험증실miR-126모의물가이촉진EPCs적천이(P＜0.01),miR-126억제물억제EPCs적천이(P＜0.01).(3)TargetScan재선연건예측KANK2시miR-126적파기인.(4) Western blot검측결과현시miR-126모의물억제KANK2적표체,miR-126억제물촉진KANK2적표체.결론 miR-126대EPCs적증식유일과성적촉진작용,miR-126가이촉진EPCs적천이능력병파향KANK2단백,억제KANK2단백적표체.
Objective To investigate the role of miR-126 (micro RNA-126) in rat endothelial progenitor 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.