驱动蛋白家族成员4A对乙肝病毒复制影响的研究

摘要

Objective To explore the effect of kinesin family member 4A protein(KIF4A) on the replication of Hepatitis B virus (HBV) and its regulatory mechanism. Methods HepG2. 2. 15 cells were transfected with different quality of plasmid carrying K1F4A gene,HBsAg and HBeAg in the supernatants were measured by ELISA,covalently closed circular DNA(ccc DNA) levels were measured by real-time PCR. HepG2 cells were co-transfected with different concentration of Flag2B-KIF4A, HBV infectious clone pHBV1. 3 and its promoter pHBV-Luc. HBV,and promoter activity were measured by luminometer fluorescence detector. Results Expression of HBsAg, HBeAg and cccDNA were upregulated by KIF4A as well as HBV promoter activity in a dose-dependent manner. Conclusion KIF4A enhances the replication of HBV in HepG2 cells.%目的 探讨驱动蛋白家族成员4A(KIF4A)对乙肝病毒(HBV)复制的影响及其调节机制.方法 将不同质量的KIF4A真核表达载体Flag2B-KIF4A转染HepG2.2.15细胞后,采用ELISA法对细胞上清液中的HBsAg和HBeAg进行定量检测；荧光定量PCR法检测细胞闭合环状DNA(cccDNA)的水平；将不同质量的Flag2B-KIF4A与HBV感染性克隆pHBV1.3、启动子pHBV-Luc共转染HepG2细胞后,采用荧光检测仪检测HBV启动子活性.结果 KIF4A能够上调细胞上清液中HBsAg、HBeAg和cccDNA的水平,并上调HBV启动子活性,且调节作用呈剂量依赖效应.结论 KIF4A在HepG2细胞可以促进HBV的复制.