SAMHD1对HBV DNA形成的抑制作用及其机制研究

发布时间：2018-10-26 18:17

【摘要】：目的:研究不同肝细胞SAMHD1表达水平及其对HBV DNA形成的抑制作用和相关机制。方法:通过实时荧光定量PCR和Western blot法检测Jurkat、THP-1、Hep G2、Hep G2.2.15和Huh7细胞等细胞SAMHD1 mRNA和蛋白表达水平;通过对Hep G2.2.15细胞转染过表达质粒和shRNA,升高或者降低SAMHD1的表达水平,观察不同SAMHD1表达水平对HBV DNA水平的影响;补充不同浓度的d NTP,比较不同d NTP浓度对HBV DNA水平的影响。结果:肝癌细胞系有较高的SAMHD1表达水平;降低SAMHD1可以使HBV DNA水平升高2.3~2.8倍,升高SAMHD1可以使HBV DNA降低至33%左右(P0.01);升高d NTP浓度可以促进HBV DNA的合成水平,抵消SAMHD1的抑制作用。结论:SAMHD1可以抑制肝癌细胞内HBV DNA的生成,可能是通过降低细胞内d NTP浓度实现的。
[Abstract]:Aim: to study the expression of SAMHD1 in different hepatocytes and its inhibitory effect on the formation of HBV DNA and its related mechanisms. Methods: the expression of SAMHD1 mRNA and protein in Jurkat,THP-1,Hep G2Hep G2.2.15 and Huh7 cells were detected by real-time quantitative PCR and Western blot. The effect of different SAMHD1 expression levels on the expression of HBV DNA was observed by transfection of SAMHD1 plasmid and shRNA, into Hep G2.2.15 cells. The effects of different d NTP concentrations on HBV DNA levels were compared by adding d NTP, at different concentrations. Results: the expression of SAMHD1 in HCC cell line was higher than that in HCC cell line, the HBV DNA level increased 2.32.8-fold by decreasing SAMHD1, and HBV DNA decreased to about 33% by increasing SAMHD1 (P0.01). Increasing d NTP concentration could promote the synthesis of HBV DNA and counteract the inhibition of SAMHD1. Conclusion: SAMHD1 can inhibit the production of HBV DNA in hepatoma cells, which may be achieved by decreasing the concentration of d NTP.
【作者单位】：浙江大学基础医学院免疫学研究所;
【分类号】：R392

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