HIV-1Vpr体外诱导肿瘤细胞凋亡与细胞周期阻滞的研究
Cell cycle arrest and cell apoptosis induced by HIV-1 Vpr in cancer cells in vitro
摘要目的 研究HIV-1 Vpr诱导Hela、Lovo、HepG2细胞凋亡和G2期阻滞效果.方法 构建含有HIV-1 Vpr基因的pcDNA4-Vpr重组载体,体外转染Hela、Lovo、HepG2细胞,同时设pcDNA4-EGFP质粒转染对照组、FUGENE组和空白对照组.转染后24、48、72 h,通过甲臜化合物法检测细胞增殖,流式细胞仪检测细胞周期分布,并使用ANNEXIN V/P1双染法检测细胞凋亡.结果 与对照组相比,转染后24、48、72 h,Hela、Lovo、HepG2三种细胞的增殖抑制率、G2期阻滞率及细胞凋亡率均呈上升趋势(P<0.05).Hela、Lovo、HepG2在转染后72 h的增殖抑制率分别为29.67%、27.35%、31.67%;G2期细胞周期阻滞率分别为24.9%、18.8%、32.1%;凋亡率分别为15.46%、7.7%、41.5%.结论 HIV Vpr体外能诱导Hela、Lovo、HepG2肿瘤细胞的G2期阻滞和细胞凋亡.
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abstractsObjective To investigate the G2 cell cycle arrest and apoptosis induced by HIV-1 Vpr gene on Hela,Lovo and HepG2 cancer cells.Methods Recombinant vector pcDNA4-Vpr was constructed by DNA recombination technology and was then transfected into Hela,Lovo and HepG2 cells.Meanwhile,pcDNA4-EGFP plasmid group,FUGENE group and blank group were also set up as control.Ratio of cytostasis was evaluated by MTS assay 24 h,48 h and 72 h later,cell cycle arrest examined by flow cytometry and apoptosis detected by staining with ANNEXIN V and PI double dyes.Results Compared to the control group,the value of inhibition ratio,G2 arrest and apoptosis of Hela,Lovo and HepG2 cells increased obviously 24 h,48 h and 72 h after the transfection ( P < 0.05 ).72 h after the transfection,the inhibition ratio of Hela,Lovo and HepG2 was 29.67%,27.35% and 31.67% respectively.Percentage of G2 phase cells was 24.9%,18.8%and 32.1% respectively.Apoptosis percentage of Hela,Lovo and HepG2 ceils was 15.46%,7.7% and 41.5% correspondingly.Conclusion HIV-1 Vpr gene can induce cell cycle G2 arrest and apoptosis of Hela,Lovo and HepG2 cell lines in vitro.
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