Inhibition of HBV DNA replication and expression in 2.2.15 hepatoma cells infected with AFP-mediated HBX antisense RNA.
- Author:
Chun-hong MA
1
;
Wen-sheng SUN
;
Su-xia LIU
;
Xiao-yan WANG
;
Li-ning ZHANG
;
Ying-lin CAO
;
Li-hui HAN
Author Information
- Publication Type:Journal Article
- MeSH: 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
- From: Chinese Journal of Hepatology 2003;11(5):291-294
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the specific expression of the antisense RNA against hepatitis B virus X (HBX) gene in hepatoblastoma cell line and its anti -HBV activity.
METHODSHBX 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.
RESULTSThe 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.
CONCLUSIONSThe pEBAF-as-HBX expression vector may lead to targeted-expression of HBX antisense RNA in hepatoma cells and shows great inhibition effect on HBV.
