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目的在Vero细胞、RD细胞和U87细胞3种细胞系中,研究EV71的2A蛋白对宿主细胞增殖能力的影响及对细胞的损伤作用,揭示2A蛋白在EV71的致病机制中所起的作用。方法将野毒株EV71SDLY 107株(强毒株)、EV71SDLY 1株(弱毒株)和嵌合毒株EV71SDLY 107-2A-1感染Vero细胞、RD细胞、U87细胞后,利用MTT试验测定3种细胞在感染病毒后不同时间点的存活情况,绘制细胞生长曲线,比较细胞增殖程度,并利用单因素方差分析法分析3种细胞在感染不同病毒48h后的存活率;结果 EV71SDLY 107株感染Vero细胞和RD细胞后细胞增殖程度和存活率均低于EV71SDLY 1株(t=41.64,P<0.001;t=33.28,P<0.001);EV71SDLY 107-2A-1嵌合株感染Vero细胞和RD细胞后,细胞增殖程度和存活率均高于亲本病毒EV71SDLY 107株,低于EV71SDLY 1株(t=17.97,P<0.001;t=42.09,P<0.01);EV71SDLY 107-2A-1株感染U87细胞后细胞存活率低于EV71SDLY 1株(t=8.85,P<0.001),但与EV71SDLY 107株感染后细胞存活率无统计学差异(t=0.74,P=0.603);不同细胞感染同株病毒48h后,U87细胞存活率均为最低,Vero细胞存活率均为最高。结论 EV71强毒株的2A蛋白对细胞产生的损伤作用大于弱毒株的2A蛋白,提示2A蛋白在EV71对宿主细胞的作用中发挥功能。
OBJECTIVE: To investigate the effect of 2A protein of EV71 on the proliferation of host cells and its damage on cells in Vero cells, RD cells and U87 cell lines to reveal the role of 2A protein in the pathogenesis of EV71. Methods Vero cells, RD cells and U87 cells were infected with EV71SDLY 107, EV71SDLY 1 and EV71SDLY 107-2A-1, respectively. MTT assay was used to determine the expression of three kinds of cells The cell growth curve was drawn and the degree of cell proliferation was compared. The survival rate of three kinds of cells after infection with different viruses for 48h was analyzed by one-way ANOVA. Results The EV71SDLY 107 strain was infected with Vero cells and The proliferation and viability of RD cells were lower than that of EV71SDLY (t = 41.64, P <0.001; t = 33.28, P <0.001). After infection of Vero cells and RD cells by EV71SDLY 107-2A- The cell proliferation rate and survival rate were higher than that of the parental virus EV71SDLY107, lower than the EV71SDLY1 strain (t = 17.97, P <0.001; t = 42.09, P <0.01) The survival rate was lower than that of EV71SDLY (t = 8.85, P <0.001), but there was no significant difference between EV71SDLY 107 and infected cells (t = 0.74, P = 0.603) U87 cell survival rate was the lowest, Vero cell survival rate was the highest. Conclusions The 2A protein of virulent strains of EV71 exerts greater effects on cell injury than the 2A protein of attenuated strains, suggesting that 2A protein functions in the function of EV71 on host cells.