title: [Inhibition of HBV DNA replication and expression in 2.2.15 hepatoma cells infected with AFP-mediated HBX antisense RNA].
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Institute of Immunology, Medical School of Shandong University, Jinan 250012, China.
authors.forename: Chun-hong
authors.initials: CH
authors.lastname: Ma
authors.forename: Wen-sheng
authors.initials: WS
authors.lastname: Sun
authors.forename: Su-xia
authors.initials: SX
authors.lastname: Liu
authors.forename: Xiao-yan
authors.initials: XY
authors.lastname: Wang
authors.forename: Li-ning
authors.initials: LN
authors.lastname: Zhang
authors.forename: Ying-lin
authors.initials: YL
authors.lastname: Cao
authors.forename: Li-hui
authors.initials: LH
authors.lastname: Han
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000473:pathology / Q000821:virology*; D045744:Cell Line, Tumor; D004261:DNA Replication; D004742:Enhancer Elements, Genetic / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D015316:Genetic Therapy / Q000379:methods; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology / Q000821:virology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D016372:RNA, Antisense / Q000494:pharmacology*; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics; D015533:Transcriptional Activation; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins; D000509:alpha-Fetoproteins / Q000235:genetics*
chemical_list: D016372:RNA, Antisense; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; D000509:alpha-Fetoproteins; C057628:hepatitis B virus X protein