Studies On Human Hepatocarcinogenesis

Aim: To investigate the hepatocarcinogenic role of HBV integration into hepatocyte genome as well as to explore a new in vitro cell system capable of expressing multiple HBV genes.Materials and Methods: 7 HBsAg sero-positive HCC surgical samples from Qidong were randomly selected for integration site studies. Extracted DNAs were subjected to Alu-PCR or "Genome Walking" techniques to amplify the DNA segments containing the insertion sites between HBV X gene region and host genome. Enriched PCR segments were DNA sequenced. The insertion sites between viral and cellular sequences were identified through homology comparison using Entrez BLAST softwares. Special emphasis was made to identify whether the HBV insertion had hit critical regions of important cellular genes.For exploring HBV expression system, a DNA recombinant was first constructed by ligating the DNA segment containing complete HBV genome into retroviral pCLXSN expression vector. After molecular...