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

发布时间：2018-10-05 08:20

【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Taq Man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SIVmac239P27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。
[Abstract]:Objective to study the relationship between the changes of gene and protein levels of SIV in CEMx174 cells and the level of virus. Methods after SIVmac239 virus was infected with CEMx174 cells, supernatants and cells were collected daily. The relative expression of SAMHD1 mRNA was detected by Taq Man probe real-time quantitative RT-PCR method, and the changes of SAMHD1 and SIVmac239P27 proteins were detected by Western blot. And analyze its correlation. Results after CEMx174 cells were infected with SIVmac239, the relative expression of SAMHD1 mRNA based on internal reference gene GAPDH increased gradually, and increased 14% and 16% respectively in the first 3 days, and then increased rapidly. The expression of SAMHD1 protein in the cells did not change at the 1st and 2nd day after infection, then decreased gradually, and was almost undetectable at the 5th and 6th day, showing a decreasing trend. Conclusion the expression of SAMHD1 gene in CEMx174 cells was up-regulated after SIVmac239 infection, which was positively correlated with the viral load in the supernatant. The intracellular SAMHD1 protein level was negatively correlated with the SIVmac239 P27 protein level.
【作者单位】：北京协和医学院比较医学中心中国医学科学院医学实验动物研究所卫生部人类疾病比较医学重点实验室国家中医药管理局人类疾病动物模型三级实验室;
【基金】：国家十二五科技重大专项课题(2013ZX10004608-003和2012ZX10004501-001)
【分类号】：R512.91

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