[Inhibition of hepatitis B virus by combined siRNAs in vitro].

Abstract

Small interfering RNA (siRNA) technology is a powerful tool to knockdown gene expression in mammalian cells including genes of viral origin. To study the inhibition of hepatitis B virus (HBV) by combined siRNAs in vitro, HepG2 2.2.15, a human hepatoblastoma cell that constitutively produces HBV particles, was transfected with 12 siRNAs that were designed to target different regions on HBV genome. Enzyme-linked immunosorbent assay was used to measure the HBsAg and HBeAg levels in culture media, and HBV replication was determined by real-time quantitative PCR. The results showed that HBV replication could be inhibited by treatment with these siRNAs in a dose-dependent manner. The expression of HBsAg was inhibited by 80% and 90% with 80 nmol/L single siRNA treatment and 20 nmol/L combined siRNAs treatment, respectively. HBeAg was inhibited by about 60% with single siRNA treatment, and the inhibition didn't increase much in the case of combined siRNAs treatment. HBV replication was inhibited by 90% with 80 nmol/L single siRNA treatment or 20 nmol/L combined siRNAs treatment. These data provide solid evidence for developing new approaches in HBV treatment with combined siRNAs.