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Chinese Journal of Hepatology ; (12): 291-294, 2003.
Artigo em Chinês | WPRIM | ID: wpr-344418

RESUMO

<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>


Assuntos
Animais , Humanos , Carcinoma Hepatocelular , Genética , Patologia , Virologia , Linhagem Celular Tumoral , Replicação do DNA , Elementos Facilitadores Genéticos , Genética , Regulação Viral da Expressão Gênica , Terapia Genética , Métodos , Vírus da Hepatite B , Genética , Fisiologia , Neoplasias Hepáticas , Genética , Patologia , Virologia , Regiões Promotoras Genéticas , Genética , RNA Antissenso , Farmacologia , Transativadores , Genética , Ativação Transcricional , Transfecção , alfa-Fetoproteínas , Genética
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