【摘要】：目的研究SIV在感染CEMx174細(xì)胞過程中,SAMHD1在基因和蛋白水平的變化與病毒水平之間的相關(guān)性。方法 SIVmac239病毒感染CEMx174細(xì)胞后,每天收集上清和細(xì)胞,應(yīng)用Taq Man探針實(shí)時(shí)熒光定量RT-PCR方法,檢測細(xì)胞內(nèi)SAMHD1 mRNA的相對表達(dá)量的變化;用Western blot檢測SAMHD1蛋白與SIVmac239P27蛋白表達(dá)量的變化,并分析其相關(guān)性。結(jié)果 CEMx174細(xì)胞感染SIVmac239后,細(xì)胞中的SAMHD1 mRNA基于內(nèi)參基因GAPDH的相對表達(dá)量逐漸增加,前3 d分別增加了14%,16%,42%,隨后開始迅速增加,第4天達(dá)到了200%,第6天更是達(dá)到了890%;細(xì)胞中SAMHD1蛋白的表達(dá)量在感染后第1、2天時(shí)變化不大,隨后逐漸降低,到第5、6天時(shí),幾乎檢測不到,呈現(xiàn)逐漸下降的趨勢。結(jié)論 CEMx174細(xì)胞感染SIVmac239后,SAMHD1基因表達(dá)上調(diào),與上清中病毒載量水平正相關(guān),細(xì)胞內(nèi)SAMHD1蛋白水平與SIVmac239 P27蛋白水平呈負(fù)相關(guān)。
[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.
【作者單位】： 北京協(xié)和醫(yī)學(xué)院比較醫(yī)學(xué)中心中國醫(yī)學(xué)科學(xué)院醫(yī)學(xué)實(shí)驗(yàn)動(dòng)物研究所衛(wèi)生部人類疾病比較醫(yī)學(xué)重點(diǎn)實(shí)驗(yàn)室國家中醫(yī)藥管理局人類疾病動(dòng)物模型三級(jí)實(shí)驗(yàn)室;
【基金】：國家十二五科技重大專項(xiàng)課題(2013ZX10004608-003和2012ZX10004501-001)
【分類號(hào)】：R512.91

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