腺病毒介导Gax基因对血清刺激后兔血管平滑肌细胞增殖和凋亡的影响  

Effects on proliferation and apoptosis of serum-induced rabbit VSMCs by adenovirus-mediated transfer of the Gax gene

作　　者：郑辉[1] 薛松[1] 连锋[1] 胡振雷[1] 汪永义[1] 

机构地区：[1]上海交通大学医学院附属仁济医院心胸外科,上海200127

出　　处：《细胞与分子免疫学杂志》2012年第1期33-36,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：上海市科委基金资助项目(08411964100)

摘　　要：目的:通过腺病毒介导人生长终止特异性同源异形盒基因(Gax)转染体外兔血管平滑肌细胞(VSMCs),观察人Gax基因过表达对血清刺激后兔VSMCs增殖和凋亡的影响,为Gax基因成为理想的静脉桥再狭窄基因治疗的候选基因提供实验基础。方法:Ad5-hGax重组腺病毒载体转染兔VSMCs后,采用Western blot检测不同时间细胞中hGax蛋白的表达;MTT法检测Ad5-hGax对血清刺激后兔VSMCs的体外增殖抑制情况;转染72 h后流式细胞术检测Ad5-hGax诱导血清刺激后VSMCs的凋亡率。结果:转染后1 d、3 d、5 d,Ad5-hGax组细胞中的hGax蛋白均有明显表达;MTT法检测显示hGax基因过表达能明显抑制血清刺激后兔VSMCs的增殖;流式细胞术检测显示转染72 h后,hGax过表达能显著诱导血清刺激后兔VSMCs的凋亡。结论:hGax基因过表达能有效抑制血清刺激后VSMCs的增殖,并促进其凋亡,为腺病毒介导Gax基因治疗静脉桥再狭窄提供重要的实验基础。AIM： To investigate the effect of a recombinant replication-incompetent AdS-hGax vector on the prolif- eration and apoptosis of serum-induced rabbit vascular smooth muscle cells （VSMCs） in vitro, and to provide an important support that Gax would be an optimal gene for gene therapy in vein graft failure. METHODS： The AdShGax vector was infected into rabbit VSMCs, then the protein level of rabbit VSMCs was detected at 1 d, 3 d, 5 d by Western blot, respectively. MTT was applied to observe the inhibitory effects of overexpressed hGax on serum-induced rabbit VSMCs. the apoptosis of serum-induced rabbit VSMCs was measured by using flow cytometry in 72 h after transfection. RESULTS： The protein expression of human Gax in the Ad5-hGax transfected cells on 1st day, 3rd day, 5th day was determined; MTT showed that the proliferation of serum-induced rabbit VSMCs was significantly inhibited in AdS-hGax group at 48 h, 72 h and 96 h （ P 〈 0.05, respectively） ; After transfecting for 72 h, flow cytometry analysis showed that the number of the apoptosis cells was in- creased in serum-induced Rabbit VSMCs of AdS-hGax group （ P 〈 0.05）. CONCLUSION： Overexpressed hGax could inhibit the proliferation of serum-induced Rabbit VSMCs and induce their apoptosis. These findings carry significant implications for adenovirus vector-based Gax gene therapies for vein graft failure.

关 键 词：生长终止特异性同源盒基因 腺病毒载体 细胞增殖 细胞凋亡 静脉桥再狭窄 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]