乙肝病毒MHBs^t/HBx蛋白对Galβ1,3GalNAcα2,3-唾液酸转移酶的反式激活作用

Regulation of Galβ1,3GalNAcα2,3-sialyltransferase (ST3GalI) by Hepatitis B Virus MHBs^t/HBx Transactivator

PCR方法扩增乙肝病毒MHBst、HBx基因片段 ,构建真核表达载体pcDNA3 1 MHBst 和pcDNA3 1 HBx。PCR方法从肝细胞基因组中扩增出Galβ1,3GalNAcα2 ,3 唾液酸转移酶 (ST3GalI)的启动子Psial,用Psial取代pEGFP N1的启动子pCMV构建pEGFP N1 Psial。利用磷酸钙 DNA共沉淀的方法 ,将pcDNA3 1 MHBst、pcDNA3 1 HBx分别与pEGFP N1 Psial瞬时共转染至正常肝细胞QGY 770 1。流式细胞仪分析细胞平均荧光密度值发现 ,MHBst、HBx分别将ST3GalI启动子的活性上调了 35 2 %和 43 8%。研究了乙肝病毒MHBst、HBx对ST3GalI的转录调控作用 。

Hepatitis B virus MHBs t and HBx fragments were amplified to construct eukaryotic expression vector pCDNA3.1-MHBs t and pCDNA3.1-HBx. ST3GalI promoter region was obtained by the method of PCR and GFP report plasmid pEGFP-N1-Psial was constructed. pCDNA3.1-MHBs t or pCDNA3.1-HBx with pEGFP-N1-Psial were transiently co-transfected into QGY-7701 cells using calcium phosphate-DNA co-precipitation, respectively. The expressions of Psial-directed GFP were analyzed by FACScalibur. It was found that MHBs t/HBx could upregulate ST3GalI promoter activity by 35.2% and 43.8%, respectively. We report the regulation of ST3GalI by MHBs t and HBx transactivators. It would be helpful to further investigate the relation between hepatitis B virus infection and sialyltransferase expression.