[Inhibition of HBV DNA replication and expression in 2.2.15 hepatoma cells infected with AFP-mediated HBX antisense RNA].

To study the specific expression of the antisense RNA against hepatitis B virus X (HBX) gene in hepatoblastoma cell line and its anti -HBV activity.
HBX gene (nt.1370-1827) was amplified by PCR, then cloned into EB virus vector pEBAF which contained human alpha-fetoprotein promoter and enhancer. After transfected into 2.2.15 hepatoma cells and ECV304 human endothelial cells by lipofectin, northern blot, ELISA and real-time qualitative PCR were carried out to assay the expression of HBX mRNA, HBV antigens and HBV DNA level, respectively.
The HBX antisense RNA expression vector pEBAF-as-HBX which could be expressed specifically in 2.2.15 hepatoblastoma cells was successfully constructed. Both HBV DNA level and the expressions of hepatitis B virus surface antigen (HBsAg) and e antigen (HBeAg) in 2.2.15 hepatoblastoma cells were inhibited by pEBAF-as-HBX. Compared with those in sense control (pEBAF-s-HBX), the inhibitory rates of HBsAg, HBeAg, and HBV DNA were 37.9%, 36.8%, and 25%, respectively.
The pEBAF-as-HBX expression vector may lead to targeted-expression of HBX antisense RNA in hepatoma cells and shows great inhibition effect on HBV.
AuthorsChun-hong Ma, Wen-sheng Sun, Su-xia Liu, Xiao-yan Wang, Li-ning Zhang, Ying-lin Cao, Li-hui Han
JournalZhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology (Zhonghua Gan Zang Bing Za Zhi) Vol. 11 Issue 5 Pg. 291-4 (May 2003) ISSN: 1007-3418 [Print] China
PMID12773245 (Publication Type: English Abstract, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • RNA, Antisense
  • Trans-Activators
  • alpha-Fetoproteins
  • hepatitis B virus X protein
  • Animals
  • Carcinoma, Hepatocellular (genetics, pathology, virology)
  • Cell Line, Tumor
  • DNA Replication
  • Enhancer Elements, Genetic (genetics)
  • Gene Expression Regulation, Viral (drug effects)
  • Genetic Therapy (methods)
  • Hepatitis B virus (genetics, physiology)
  • Humans
  • Liver Neoplasms (genetics, pathology, virology)
  • Promoter Regions, Genetic (genetics)
  • RNA, Antisense (pharmacology)
  • Trans-Activators (biosynthesis, genetics)
  • Transcriptional Activation
  • Transfection
  • alpha-Fetoproteins (genetics)

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