Inhibition of HBV DNA replication and expression in 2.2.15 hepatoma cells infected with AFP-mediated HBX antisense RNA / 中华肝脏病杂志
Chinese Journal of Hepatology
; (12): 291-294, 2003.
Article
in Zh
| WPRIM
| ID: wpr-344418
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>
Full text:
1
Database:
WPRIM
Main subject:
Pathology
/
Pharmacology
/
Physiology
/
Virology
/
Transfection
/
Genetic Therapy
/
Alpha-Fetoproteins
/
Gene Expression Regulation, Viral
/
Transcriptional Activation
/
Trans-Activators
Type of study:
Qualitative_research
Limits:
Animals
/
Humans
Language:
Zh
Journal:
Chinese Journal of Hepatology
Year:
2003
Document type:
Article