miR-155对HBV蛋白表达的抑制作用及机制

发布时间：2018-12-14 02:44

【摘要】：目的观察mi R-155对SOCS1 m RNA及蛋白水平的影响,探究mi R-155对HBs Ag、HBe Ag表达的抑制作用。方法从Hep G2.2.15细胞中提取基因组,经PCR扩增得到mi R-155前体序列,纯化并连接入pm R-m Cherry载体,构建重组质粒pmi R-155,经去内毒素后转染至Hep G2.2.15细胞。将细胞分为重组组(转染pmi R-155质粒)、空载组(转染pm Rm Cherry质粒)、转染试剂组、空白组。采用荧光实时定量PCR检测各组细胞内mi R-155的表达,RT-PCR检测各组细胞SOCS1 m RNA的表达,Western blotting检测各组细胞SOCS1 蛋白的表达,ELISA检测各组细胞HBs Ag、HBe Ag的表达。结果荧光实时定量PCR结果显示,以空白组细胞内mi R-155的表达量为基准,重组组mi R-155表达量(519.43±52.10)明显高于空载组(24.24±16.70)及转染试剂组(35.04±26.09,P0.05);RT-PCR结果显示,重组组SOCS1 m RNA表达量(0.63±0.91)显著低于空白组(P0.05);Western blotting结果显示,重组组SOCS1蛋白表达量显著低于空白组。以空白组的表达量为基准,重组组HBs Ag和HBe Ag蛋白表达的抑制率分别为55.62%±3.77%和47.87%±2.46%(P0.01)。结论过表达的mi R-155可抑制细胞内SOCS1和HBV蛋白的表达。
[Abstract]:Objective to observe the effect of mi R-155 on the level of SOCS1 m RNA and protein, and to explore the inhibitory effect of mi R-155 on the expression of HBs Ag,HBe Ag. Methods the genome was extracted from Hep G2.2.15 cells, and the mi R-155 precursor sequence was amplified by PCR, then purified and ligated into pm R-m Cherry vector. The recombinant plasmid pmi R-155 was constructed and transfected into Hep G2.2.15 cells after endotoxin removal. The cells were divided into recombinant group (transfected with pmi R-155 plasmid), no-load group (transfected with pm Rm Cherry plasmid), transfection reagent group and blank group. Fluorescence real-time quantitative PCR was used to detect the expression of mi R-155, RT-PCR to detect the expression of SOCS1 m RNA, Western blotting to detect the expression of SOCS1 protein in each group, and ELISA to detect the expression of HBs Ag,HBe Ag in each group. Results the results of real-time fluorescence quantitative PCR showed that the expression of mi R-155 was significantly higher in the recombinant group (519.43 卤52.10) than in the no-load group (24.24 卤16.70) and the transfection reagent group (35.04 卤26.09). P0.05); RT-PCR results showed that the expression of SOCS1 m RNA in the recombinant group (0.63 卤0.91) was significantly lower than that in the control group (P0.05); Western blotting). The expression of SOCS1 protein in the recombinant group was significantly lower than that in the blank group. The inhibition rates of HBs Ag and HBe Ag protein expression in the recombinant group were 55.62% 卤3.77% and 47.87% 卤2.46% respectively (P0.01). Conclusion overexpression of mi R-155 can inhibit the expression of SOCS1 and HBV proteins.
【作者单位】：广州军区广州总医院儿科;
【基金】：广州市科技计划项目(2013J4100116)~~
【分类号】：R512.62

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