反义mTOR逆转录病毒载体对血管平滑肌细胞增殖的抑制作用

目的：观察反义雷帕霉素靶蛋白(mTOR)逆转录病毒载体对血管平滑肌细胞(VSMC)增殖活性的影响。方法：采用脂质体介导转染包装细胞后构建mTOR的反义重组逆转录病毒载体(pLXIN-AmTOR)感染VSMC，检测mTOR及其下游底物包括真核细胞启动子4E结合蛋白1(4E-BP1)和核糖体蛋白S6激酶(p70S6k)的表达变化，以及VSMC细胞增殖周期和增殖活性的改变。 结果：反义逆转录病毒载体转染PT67细胞后感染VSMC，能显著抑制mTOR的mRNA和蛋白产物表达，mTOR通路下游p70S6k表达相应减少，而4E-BP1的表达却显著增强;感染后的VSMC的分裂、增殖过程受阻，更多细胞停滞在G1期。 结论：成功构建反义mTOR逆转录病毒载体;感染VSMC后能明显抑制VSMC的分裂、分化和增殖。

The inhibition of antisense RNA retroviral vector of mTOR on proliferation of vascular smooth muscle cells

Objective To study the effect of antisense RNA retroviral vector of mTOR on the proliferation of vascular smooth muscle cells(VSMC).Methods The conservative region of mTOR gene was inserted into pLXIN reversedly,then the vector was packaged in PT67 cells by transfection with lipofectamine and(transfected) to VSMC.The efficiency of antisense inhibition was verified,and the changes of p70S6k and(4E-BP1) were also determined at the same time.The proliferation activity was determined by flowcytometry and MTT.Results After the vector was successfully transfected to the cells.pLXIN-A-mTOR vector could(efficiently) decrease mRNA and protein expression of mTOR and p70S6k of the cell,while the expression of 4E-BP1 increased significantly.The cell cycle and proliferation activity of VSMC was also stunned in phase G_1.Conclusions The mTOR antisense retroviral vector,pLXIN-A-mTOR,has been constructed(successfully),which can significantly inhibit the proliferation of VSMC.