南京医科大学学报（自然科学版）2011，Vol.31Issue(2)：131-137,7.
重组慢病毒载体介导HIV-1Nef蛋白对原发性渗出性淋巴瘤细胞系和血管内皮细胞增殖作用影响的初探
Role of HIV-1 Nef protein carried by recombinant lentivirus in proliferation of primary effusion lymphoma cell line and vascular endothelial cells
摘要
Abstract
Objective: To construct the recombinant lentivirus containing HIV-1 Nef gene and detect the protein expression in its target cell lines, such as 293T, BCBL-1 and EA.hy926, and explore the effect of Nef on cell proliferation. Methods:The fragment of Nef gene from expression vector pCl-neo-Nef was cloned into the lentivirus vector pHAGE-CMV-MCS-Izs-Green, then the recombinant vector pHAGE-Nef, packaging vector psPAX2 and envelope vector pMD2.G were cotransfected into the 293T cells. Culture media were harvested and filtered through a 0.45 μm filter to remove the cells. The viral titer was checked by observing the expression of green fluorescent protein (GFP). After infection of the recombinant lentivirus, the protein expression of Nef in 293T, BCBL-1 and EA.hy926 cells were detected by Western blot. The effect of Nef on cell proliferation was investigated by Cell Counting Kit-8 assay.Results:The recombinant lentivirus vector carrying HIV-1 Nef was constructed successfully. The viral titer was l×l07 TU/ml. 293T,BCBL-1 and EA.hy926 cells could be efficiently infected by it. The expression of Nef in these cell lines could be detected by Western blot. In addition, Nef protein could inhibit the proliferation of BCBL-1 and EA.hy926 cells. Conclusion: Recombinant lentivirus with high titer and efficient infection of 293T, BCBL-1 and EA.hy926 cells, could be obtained quickly and simply by using the lentivirus vectors system, and Nef expression could be detected in 293T, BCBL-1 and EA.hy926 cells infected by lentivirus-Nef. Furthermore,our study suggested that Nef protein could inhibit the proliferation of BCBL-1 and EA.hy926 cells.关键词
慢病毒/Nef/卡波济肉瘤/卡波济肉瘤相关疱诊病毒Key words
recombinant lentivirus/ Nef/ KS/ KSHV分类
医药卫生引用本文复制引用
朱小飞,秦娣,周峰,卫冰冰,卢春..重组慢病毒载体介导HIV-1Nef蛋白对原发性渗出性淋巴瘤细胞系和血管内皮细胞增殖作用影响的初探[J].南京医科大学学报（自然科学版）,2011,31(2):131-137,7.基金项目
国家自然科学基金(30900064) （30900064）
教育部高等学校博士点专项科研基金新教师基金(20093234120004) （20093234120004）
江苏省高校自然科学基金(09KJB310007) （09KJB310007）
