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目的研究La蛋白与乙型肝炎病毒(HBV)mRNA稳定性和蛋白质表达之间的相关性。方法设计并合成La蛋白mRNA特异性小干扰RNA(siRNA),转染HepG2.2.15细胞,继续培养3d后分别裂解转染和未转染siRNA的细胞;抽提蛋白质,Western blot检测La蛋白含量变化;于转染后的第1、2、3、5、7、9天分别收集细胞培养上清液,实时荧光定量聚合酶链反应技术检测上清液中HBV DNA的变化情况,电化学发光分析技术检测乙型肝炎表面抗原,乙型肝炎e抗原含量变化情况。结果La蛋白在转染siRNA的HepG2.2.15细胞中表达明显低于未转染siRNA的细胞;转染siRNA的细胞分泌到上清液中乙型肝炎表面抗原、乙型肝炎e抗原和HBV DNA的含量也明显低于未转染siRNA的细胞。结论在HepG2.2.15细胞内,La蛋白与HBV mRNA稳定性和蛋白质表达具有功能相关性。
Objective To study the relationship between the stability of La protein and hepatitis B virus (HBV) mRNA and protein expression. Methods La mRNA was designed and synthesized. The HepG2.2.15 cells were transfected into HepG2.2.15 cells, and the cells transfected and not transfected with siRNA were further cultured for 3 days. The proteins were extracted and the La protein The cell culture supernatants were collected on the 1st, 2nd, 3rd, 5th, 7th and 9th day after transfection. The changes of HBV DNA in the supernatant were detected by real-time fluorescence quantitative polymerase chain reaction Luminescence analysis of hepatitis B surface antigen, hepatitis B e antigen content changes. Results The expression of La protein in HepG2.2.15 cells transfected with siRNA was significantly lower than that in untransfected siRNAs. The cells transfected with siRNA secreted the supernatant of hepatitis B surface antigen, hepatitis B e antigen and HBV The content of DNA was also significantly lower than that of untransfected siRNA. Conclusion In HepG2.2.15 cells, La protein has a functional relationship with HBV mRNA stability and protein expression.