FUNCTIONAL ASPECTS OF HEPATITIS B VIRUS X-PROTEIN

研究了肿瘤抑制基因Rb的瞬时表达。 HBVX基因导入Hep G2细胞表达系统的研究乙肝病毒 X开放阅读框是从含有野生型基因的质粒中通过聚合酶链式反应产生的 型HBVdna，并插入表达载体中。表达载体pHBVX 并将其应用于Hep G2细胞系中。提取的级别 用Rb单抗进行WB分析内源性Rb蛋白。 未转Rb蛋白的Hep G2细胞及载体转染的Hep G2细胞 单独的也被分析为对照。PHBVX中Rb蛋白的表达水平 与对照组相比，转染组Hep G2细胞数增加近10倍 细胞。Rb体外免疫共沉淀的初步研究 蛋白质和HBVX蛋白不表明Rb与Rb之间存在直接相互作用 和HBVX蛋白。发现乙肝病毒衍生的X基因可以改变 Rb抑癌基因的表达提示其可能在肿瘤发生、发展中的作用。 X基因在乙肝病毒介导的肝癌发生中的作用可能的作用机制 我们将探索如何激活。

The expression of tumor suppressor gene RB was studied in a transient expression system by transfecting HBV X gene into Hep G2 cells. The HBV X open reading frame was generated by PCR from a plasmid containing wild type HBV DNA and was inserted into expression vector. The plasmid pHBV-X was used to transfect a Hep G2 cell line. The level of extracted endogenous RB protein was analyzed by WB using RB monoclonal antibody. RB protein untransfected Hep G2 cells and Hep G2 transfected with vector alone were also analyzed as controls. The level of RB protein in pHBV-X transfected Hep G2 cells increased nearly 10-fold compared to the control cells. Preliminary in vitro co-immunoprecipitation studies between RB protein and HBV X protein do not indicate direct interaction between RB and HBV X proteins. The finding that HBV derived X gene can alter the expression of RB tumor suppressor gene suggest a possible role for the X gene in HBV mediated hepatocarcinogenesis. Possible mechanisms of transactivation will be explored.