90 K／Mac-2 BP 基因沉默增强 HIV-1感染的单核巨噬细胞凋亡

Gene silencing of 90K /Mac-2BP enhances the apoptosis of U937 cells by HIV-1 infection

目的研究人单核巨噬细胞株U937的90K/Mac-2BP基因沉默后其mRNA和蛋白水平的表达及其对HIV-1感染单核巨噬细胞凋亡的影响。方法用人单核巨噬细胞株U937作为细胞模型,用R5嗜性的HIV-1全长感染性质粒包装并获得HIV-1病毒,待HIV-1感染U937细胞5 d后,细胞经PE-Annexin V,PerCP-7-AAD染色,用流式细胞仪检测细胞凋亡情况。采用基因沉默技术,将U937细胞中的90K/Mac-2BP基因沉默后,再经HIV-1感染,5 d后检测细胞凋亡情况。结果正常人单核巨噬细胞株U937,经HIV-1感染后细胞凋亡率为(16.27±0.30)%,而90K/Mac-2BP基因沉默后的U937细胞,经HIV-1感染后细胞凋亡率分别增加至(31.26±0.35)%、(25.76±0.30)%、(23.69±0.33)%,具有统计学差异(P<0.05)。结论 90K/Mac-2BP的表达水平能调节HIV-1感染的单核巨噬细胞凋亡。

Objective To investigate the effect of cell death by HIV-1 infection on gene 90K/Mac-2BP by RNA interference （RNAi） in U937 cell line.Methods We used human monocyte-macrophage cell line U937 as the cell model.Cells were infected by HIV-1 （ R5-tropic） 5 days, and then stained by PE-Annexin V and PerCP-7-AAD.90K/Mac-2BP in U937 cell line was knocked down , and these cells were infected by HIV-1 for 5 days.Then, cells were stained by PE-AnnexinV and PerCP-7-AAD.Apoptosis were examined upon flow cytometry .Results The percentages of Annexin V＋cells without 90K/Mac-2BP knock-down were （16.27 ±0.30）% by HIV-1 infection.The percentages of them with 90K/Mac-2BP knock-down were （31.26 ±0.35）%, （25.76 ±0.30）%, （23.69 ±0.33）% respectively.The increase＆amp;nbsp;of cell apoptosis rate for HIV-1-infected U937 cells by 90K/Mac-2BP siRNA transfection was significantly greater than that for HIV-1-infected untreated cells （P﹤0.01）.Conclusion The apoptosis of HIV-1-infected U937 cells was regulated by the expression of 90K/Mac-2BP.