乙型肝炎病毒X基因siRNA对HepG2.2.15细胞的影响

发布时间：2018-07-24 11:00

【摘要】：目的:利用化学合成的小干扰RNA(si RNA)转染HepG2.2.15细胞,构建针对乙型肝炎病毒(HBV)X基因的细胞干扰模型,研究其在体外对HBV复制和抗原表达的抑制作用。方法 :将构建成功的HepG2.2.15细胞干扰模型,于转染后24、48、72 h,化学发光免疫法检测细胞上清液中HBsAg、HBeAg,ELISA法检测细胞上清液HBxAg蛋白表达量,荧光定量PCR检测转染后细胞中HBx mRNA相对表达量,CCK-8法检测细胞增殖能力。结果:HBx-siRNA转染HepG2.2.15细胞后,细胞增殖能力被抑制,细胞中HBx mRNA及上清液中HBxAg的表达量下降(P0.05);并发现其能抑制细胞HBsAg、HBeAg表达,抑制高峰在72 h,抑制率分别是66%和58%;荧光定量PCR证实细胞上清液中HBV DNA的表达量下降。结论:成功地构建了HBV X基因的Hep G2.2.15细胞干扰模型,在体外具有抑制Hep G2.2.15细胞增殖和HBV基因的复制与表达的作用。
[Abstract]:Aim: to construct a cell interference model for hepatitis B virus (HBV) (HBV) X gene by transfection of HepG2.2.15 cells with chemically synthesized small interfering RNA (si RNA), and to study its inhibitory effect on HBV replication and antigen expression in vitro. Methods: the successfully constructed HepG2.2.15 cell interference model was used to detect the expression of HBxAg protein in the supernatant by chemiluminescent immunoassay at 24 ~ 4872 h after transfection. Fluorescence quantitative PCR was used to detect the relative expression of HBx mRNA in transfected cells and CCK-8 method was used to detect cell proliferation. Results the proliferation of HepG2.2.15 cells was inhibited and the expression of HBxAg in HBx mRNA and supernatant was decreased (P0.05), and it was found that it could inhibit the expression of HBeAg in HepG2.2.15 cells. The inhibitory rate was 66% and 58% respectively at 72 h, and fluorescence quantitative PCR confirmed the decrease of HBV DNA expression in the supernatant. Conclusion: the interference model of Hep G2.2.15 cells with HBV X gene has been successfully constructed, which can inhibit the proliferation of Hep G2.2.15 cells and the replication and expression of HBV gene in vitro.
【作者单位】：广州医科大学附属第二医院检验科;广东省医学科学院
【基金】：广州市医药卫生科技项目(编号:20141A011082)
【分类号】：R512.62

【相似文献】