CREG活化ILK/VEGF促进内皮细胞血管新生  

Cellular Repressor of E1A-stimulated Gene on Endothelial Cell Angiogenesis Via ILK/VEGF Pathway

作　　者：闫承慧[1] 孙鸣宇[1] 张剑[1] 田孝祥[1] 陶杰[1] 张效林[1] 韩雅玲[1] 

机构地区：[1]沈阳军区总医院全军心血管病研究所全军重点实验室,辽宁沈阳110840

出　　处：《现代生物医学进展》2012年第36期7021-7025,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81070097;30971218);辽宁省科技攻关项目(200925009-9)

摘　　要：目的:探讨E1A激活基因阻遏子(Cellular repressor of E1A-stimulated gene,CREG)对血管内皮细胞血管新生的影响及其机制。方法:将重组腺病毒CREG载体(Ad-GFP-CREG)和腺病毒GFP载体(Ad-GFP)转染人血管内皮细胞株(vascularendothelialcell,VE),转染后细胞分别命名为ADCREG和ADGFP。内皮细胞基质胶血管形成实验观察其比较3组细胞血管发生情况。应用Western blotting测定细胞中CREG、整合素连接激酶(integrin-linked kinase,ILK)和血管内皮生长因子(vascular endothelial growthfactor,VEGF)的表达。应用ILK激酶活性突变的质粒转染和不同浓度的VEGF中和抗体转染VE后进行细胞血管形成能力检测。结果:体内基质胶血管形成实验显示,腺病毒介导的CREG过表达显著促进VE的血管形成能力,血管密度、数量和交叉点均较对照组ADGFP和VE显著增多(P<0.01)。同时,Westernblot分析证实CREG过表达显著增加了ILK与VEGF的表达;转染ILK激酶失活质粒导致ADCREG成血管能力显著减弱(P<0.001)。应用VEGF中和抗体干预后,ADCREG细胞的成血管能力也呈现剂量依赖性下降。并且ILK激酶活性受到抑制的ADCREG细胞中VEGF表达降低。结论:CREG基因过表达通过活化ILK/VEGF信号通路促进了内皮细胞血管新生。Objective：To evaluate the effects and mechanisms of cellular repressor of E1A-stimulated genes（CREG） on endothelial cells（ECs） angiogenesis.Methods：（1） Recombinant advirus CREG vector（Ad-GFP-CREG） and advirus GFP vector（Ad-GFP） transfected VEs and the transfected cells named as ADCREG and ADGFP;ADCREG,ADGFP and VE cells angiogenesis were investigated by seeding the cells into 24 well plate coated with matrigel;Western blotting was used to detected CREG,integrin-linked kinase（ILK） and vascular endothelial growth factor（VEGF） in ADCREG and ADGFP cells.ILK plasmid with kinase-dead mutant was transfected into VE cells and G418 was used to select stably-transfected cells.Matrigel angiogenesis assay was used to measure the angiogenesis ability of ILK-plasmid-transfecting ADCREG;The effect of VEGF neutralizing antibody（VEGF NA） on ADCREG angiogenesis also be assessed.Results：CREG ovexpression induced VE angiogenesis accompanied by up-regulation of ILK and VEGF expression.The unbinding of ILK kinase dead weakened significantly ADCREG angiogenesis（P 0.001）;VEGF165 NA inhibited ADCREG angiogenesis in a dose-dependent manner.Conclusions：CREG induced angiogenesis by up-regulating ILK/VEG F165 expression.

关 键 词：E1A蛋白(E1A)阻遏子 平滑肌细胞 迁移 

分 类 号：Q593[生物学—生物化学] Q75