整合素β1在CD151促HUVEC迁移增殖中的机制研究

MECHANISM OF INTEGRIN β1 IN CD151 TO PROMOTE THE PROLIFERATION AND MIGRATION OF HUVEC

目的研究整合素β1在CD151促脐静脉内皮细胞迁移增殖中的机制。方法通过构建PAAV-CD151质粒及其pAAV-CD151-AAA194-196突变体(QRD)并转染HUVEC,HUVEC分为正常对照组、绿色荧光组(GFP组)、CD151组,QRD组。SRB法检测细胞的增殖,划痕试验检测细胞的迁移能力,Western blot检测CD151、Akt、P-Akt、PI3K及β1的表达。结果1、CD151组与对照组及GFP组相比具有明显促进细胞增殖的能力,而QRD组则显著抑制细胞的增殖,具有显著的统计学意义(P<0.05)。2、CD151组与对照组及GFP组相比具有明显促进细胞迁移的能力,而QRD组则显著抑制细胞的迁移,具有显著的统计学意义(P<0.05)。3、CD151组β1、PI3K、P-Akt蛋白表达量明显增高,与对照组、GFP组和QRD组相比具有显著差异(P<0.05),QRD组蛋白表达量明显降低具有显著差异(P<0.05),而总的Akt四组之间无明显差异(P>0.05)。结论CD151具有促进血管形成的作用,通过整合素β1上调PI3K的表达,进一步促进Akt的磷酸化,增加Akt的活力,达到促进内皮细胞的迁移和增殖及管状结构的形成,进而促进血管形成。

Objective To study the mechanism of integrin β1 in CD151 to promote the proliferation and migration of human umbilical vein endothelial cells.Methods We constructed pAAV-CD151-AAA^194-196 mutant（QRD） and transfected it into HUVEC.HUVEC were divided into the normal control group,group of green fluorescence（GFP group）,CD151 group,and QRD group.The proliferation activity of HUVEC was measured by SRB assay,the ability of cell migration was detected by scratch test,and the expressions of CD151,Akt,P-Akt,PI3K,and β1 were tested by Western blot.Results 1.The CD151 group,compared with the control and GFP groups,significantly promoted cell proliferation,while the QRD group significantly inhibited cell proliferation,with significant statistical differences（P〈0.05）.2.The CD151 groups,compared with the control and GFP groups,significantly promoted cell migration,while the QRD group significantly inhibited cell migration,with significant statistical differences（P〈0.05）.3.The expressions of CD151,P-Akt,PI3K,and β1 in the CD151 group were significantly increased compared with those of the control,GFP and QRD groups（P〈0.05）,while the expression of total AKT in the four groups had no statistical significance（P〈0.05）.Conclusion CD151 has a role in promoting angiogenesis by increasing the expression of PI3K through integrin β1,to further promote the phosphorylation of Akt and increase the vitality of Akt,to promote endothelial cell migration and proliferation,and to promote the formation of tubular structures,thus contributing to angiogenesis.