FOXO3a过表达对内皮祖细胞周期相关蛋白的调控

目的:观察FOXO3a(forkhead box O3a)的活性改变对内皮祖细胞(endothelial progenitor cells,EPCs)增殖和细胞周期相关蛋白表达的影响。方法:将携带突变激活FOXO3a基因的腺病毒载体Ad-TM(triple mutant)-FOXO3a和阴性对照腺病毒载体Ad-GFP体外感染人脐血来源的EPCs。观察EPCs形态学改变,CCK-8分析转染后EPCs增殖情况,Western blot检测FOXO3a蛋白、细胞周期相关蛋白p27kip1以及CDK2的表达水平。结果:构建了的2种腺病毒相关载体被成功转染。形态学改变方面,Ad-TM-FOXO3a组EPCs细胞生长缓慢,集落不明显;Western blot和CCK-8结果显示,Ad-TM-FOXO3a转染组与阴性对照组相比,EPCs增殖被抑制,FOXO3a与p27kip1蛋白过表达,CDK2表达下调。结论:FOXO3a可能通过上调p27kip1蛋白表达,下调CDK2表达,以抑制EPCs增殖。

The Effects of FOXO3a Overexpression on the Cell Cycle RegulatoryProteins in Endothelial Progenitor Cells

Objective:To investigate the effects of FOXO3a overexpression on the proliferation of endothelial progenitor cells(EPCs) and the expression of the cell cycle regulatory proteins.Methods:Two constructed recombinant adenovirus vectors, Ad-TM(triple mutant)-FOXO3a and the control Ad-GFP, were transfected into EPCs from human cord blood in vitro. The changes in cellmorphology and proliferation of transfected EPCs were detected by fluorescent microscope and CCK-8 assay, respectively. The proteinexpression changes of FOXO3a, cell cycle regulatory proteins p27kip1 and CDK2 were assessed by Western blot.Results:The bothrecombinant adenovirus vectors were successfully transfected into EPCs. The results demonstrated that compared with control group, inthe Ad-TM-FOXO3a group had the overexpressed-FOXO3a, while cell division proceeded slowly and proliferation EPCs wassignificantly inhibited; Meanwhile p27kip1 were upregulated while CDK2 expression was downregulated.Conclusion:FOXO3a mayinhibit proliferation of EPCs via up-regulating the expression of p27kip1 and down-regulating the expression of CDK2.