腺病毒介导gax基因转染在培养的血管平滑肌细胞中的表达  

The expression of gax gene in cultured vascular smooth muscle cells mediated by adenovirus transfection

作　　者：王耿[1] 何国祥[1] 张萍[1] 冉擘力[1] 刘建平[1] 陈渝[1] 

机构地区：[1]第三军医大学附属西南医院心血管内科重庆市介入心脏病学研究所,400038

出　　处：《介入放射学杂志》2003年第S1期75-77,共3页Journal of Interventional Radiology

摘　　要：目的　以腺病毒介导gax基因转染血管平滑肌细胞 (VSMC) ,增强VSMC中gax基因的表达。方法　采用位置特异性重组方法构建携带大鼠 gax基因表达序列的复制缺陷型 5型腺病毒载体(AdCMV gax) ,经 2 93细胞扩增 ,纯化制备高滴度病毒转染液 ;以病毒转染液常规转染VSMC后 ,应用RT PCR、流式细胞仪和免疫细胞化学等方法分别检测VSMC中 gaxmRNA和蛋白质的表达。 结果　流式细胞仪检测和免疫细胞化学染色均显示AdCMV gax转染VSMC后 ,VSMC的Gax蛋白表达率明显增高 ,转染后 2 4小时即可达 80 %左右 ,高水平的表达可维持 5天以上 ;AdCMV gax转染前 ,PDGF BB对gax基因转录和翻译水平的表达均有下调作用 ,AdCMV gax转染后 ,无论有无PDGF BB刺激 ,VSMC中 gax基因的表达均比转染前显著增高。 结论　腺病毒载体可有效地介导 gax基因转染VSMC ,并且表达为蛋白质。这有利于进一步研究Objective Overexpressing gax gene in vascular smooth muscle cells(VSMC) mediated by adenovirus transfection.Methods Replication defective adenovirus type 5 (Ad5) vector encoding the rat gax gene (AdCMV gax) was constructed via site specific recombination and amplified in 293 cells. The resulting adenovirus was purified for a high viral titer. The expression of gax mRNA and protein was evaluated by a series of methods such as reverse transcription polymerase chain reaction (RT PCR), flow cytometry ...

关 键 词：GAX基因 血管平滑肌细胞 腺病毒 

分 类 号：R346[医药卫生—基础医学]