CEMx174细胞中 SAMHD1蛋白表达与SIVmac239病毒水平的相关性研究

摘要

目的：研究SIV在感染CEMx174细胞过程中，SAMHD1在基因和蛋白水平的变化与病毒水平之间的相关性。方法 SIVmac239病毒感染CEMx174细胞后，每天收集上清和细胞，应用TaqMan探针实时荧光定量RT-PCR方法，检测细胞内SAMHD1 mRNA的相对表达量的变化；用Western blot检测SAMHD1蛋白与SIVmac239 P27蛋白表达量的变化，并分析其相关性。结果 CEMx174细胞感染SIVmac239后，细胞中的SAMHD1 mRNA基于内参基因GAPDH的相对表达量逐渐增加，前3 d分别增加了14％，16％，42％，随后开始迅速增加，第4天达到了200％，第6天更是达到了890％；细胞中SAMHD1蛋白的表达量在感染后第1、2天时变化不大，随后逐渐降低，到第5、6天时，几乎检测不到，呈现逐渐下降的趋势。结论 CEMx174细胞感染SIVmac239后，SAMHD1基因表达上调，与上清中病毒载量水平正相关，细胞内SAMHD1蛋白水平与SIVmac239 P27蛋白水平呈负相关。%Objective To explore the relationship between protein levels of SAMHD1 expression with viral load of SIV in the SIVmac239-infected CEMx174 cells.Methods CEMx174 cells were infected with SIVmac239, and the cells and supernatant were collected daily for analysis.The viral loads and the SAMHD1 mRNA level were detected by real-time RT-PCR and the level of P27 and SAMHD1 proteins were detected by Western blotting.Results The mRNA level of intracellular SAMHD1 gene was increased and SAMHD1 protein decreased gradually when CEMx174 cells were infected with SIVmac239.Conclusions There is a positive correlation between viral load in the supernatant and the mRNA level of SAMHD1, and a negative correlation between the increase of P27 protein and the decrease of SAMHD1 protein in the SIVmac239-infected CEMx174 cells.