论文部分内容阅读
探讨HIV-1感染宿主细胞后对其宿主蛋白肿瘤易感基因101蛋白(Tumor Susceptibility Gene 101,TSG101)及ALG-2相互作用蛋白X(ALG-2-interacting protein X,Alix)表达的影响。以HIV-1感染性克隆病毒pNL4-3感染TZM-bl PM1、Jurkat细胞株和人外周血单个核细胞(PBMCs),感染24h后收获细胞提取总RNA,逆转录PCR检测在RNA水平各因子的表达差异;感染48h后收获细胞提取总蛋白,Western-blot检测各因子在蛋白水平的表达差异。结果显示:HIV-1感染对原代PBMC与细胞系表达Alix与TSG101影响显著不同,细胞系主要表现为下调,而原代PBMC主要表现为TSG101上调;细胞系中的下调又细分为Jurkat细胞的Alix与TSG101的双下调、TZM-bl细胞的Alix单下调以及PM1细胞无影响三种情况。HIV-1感染对细胞宿主分子TSG101及Alix在RNA和蛋白水平的表达均有影响,这种影响因细胞的不同而有差异。HIV-1感染调节Alix与TSG101的机制生物学意义尚有待于进一步阐明。
To investigate the effect of HIV-1 infection on the expression of Tumor Susceptibility Gene 101 (TSG101) and ALG-2-interacting protein X (Alix) in host cells infected by HIV-1. TZM-bl PM1, Jurkat cells and human peripheral blood mononuclear cells (PBMCs) were infected with HIV-1 infectious clone virus pNL4-3, and the total RNA was harvested 24h after infection. Reverse transcriptase PCR was used to detect the levels of various factors After 48 hours of infection, the cells were harvested to extract the total protein. Western-blot was used to detect the difference in protein expression. The results showed that: HIV-1 infection significantly affected the expression of Alix and TSG101 in primary PBMCs and cell lines, while the primary ones were down-regulated while the primary PBMCs were up-regulated by TSG101. The down-regulation in cell lines was subdivided into Jurkat cells Alix and TSG101 dual down regulation, Alix single down regulation of TZM-bl cells and PM1 cells had no effect on three cases. HIV-1 infection affects the expression of cellular host molecules TSG101 and Alix both at the RNA and protein levels, and this effect varies from cell to cell. The biological significance of the mechanisms regulating Alix and TSG101 by HIV-1 infection remains to be elucidated.