FOX03a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p27^kip1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；Westem 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p27^kip1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 Regulatory Proteins 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 cell morphology and proliferation of transfected EPCs were detected by fluorescent microscope and CCK-8 assay, respectively. The protein expression changes of FOXO3a, cell cycle regulatory proteins p27kipl and CDK2 were assessed by Western blot. Results： The both recombinant adenovirus vectors were successfully transfeeted into EPCs. The results demonstrated that compared with control group, in the Ad-TM-FOXO3a group had the overexpressed-FOXO3a, while cell division proceeded slowly and proliferation EPCs was significantly inhibited; Meanwhile p27kipl were upregulated while CDK2 expression was downregulated. Conclusion： FOXO3a may inhibit proliferation of EPCs via up-regulating the expression ofp27kipl and down-regulating the expression of CDK2.