Inhibition of hepatitis B virus gene expression and replication by artificial microRNA

World J Gastroenterol. 2008 Aug 7;14(29):4684-9. doi: 10.3748/wjg.14.4684.


Aim: To investigate the inhibitory effects of hepatitis B virus (HBV) replication and expression by transfecting artificial microRNA (amiRNA) into HepG2.2.15 cells.

Methods: Three amiRNA-HBV plasmids were constructed and transfected into HepG2.2.15 cells. HBV antigen secretion was detected in the cells with transient and stable transfection by time-resolved fluoroimmunoassays (TRFIA). HBV DNA replication was examined by fluorescence quantitative PCR, and the level of HBV S mRNA was measured by semi-quantitative RT-PCR.

Results: The efficiency of transient transfection of the vectors into 2.2.15 cells was 55%-60%. All the vectors had significant inhibition effects on HBsAg and HBeAg at 72 h and 96 h after transfection (P < 0.01 for all). The secretion of HBsAg and HBeAg into the supernatant was inhibited by 49.8% +/- 4.7% and 39.9% +/- 6.7%, respectively, at 72 h in amiRNA-HBV-S608 plasmid transfection group. The copy of HBV DNA within culture supernatant was also significantly decreased at 72 h and 96 h after transfection (P < 0.01 for all). In the cells with stable transfection, the secretion of HBsAg and HBeAg into the supernatant was significantly inhibited in all three transfection groups (P < 0.01 for all, vs negative control). The copies of HBV DNA were inhibited by 33.4% +/- 3.0%, 60.8% +/- 2.3% and 70.1% +/- 3.3%, respectively.

Conclusion: In HepG2.2.15 cells, HBV replication and expression could be inhibited by artificial microRNA targeting the HBV S coding region. Vector-based artificial microRNA could be a promising therapeutic approach for chronic HBV infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carcinoma, Hepatocellular / metabolism
  • Carcinoma, Hepatocellular / pathology
  • Carcinoma, Hepatocellular / virology
  • Cell Line, Tumor
  • DNA Replication / drug effects
  • DNA, Viral / genetics
  • Gene Expression Regulation, Viral / drug effects*
  • Hepatitis B Surface Antigens / metabolism
  • Hepatitis B e Antigens / metabolism
  • Hepatitis B virus / genetics*
  • Hepatitis B virus / immunology
  • Hepatitis B virus / physiology*
  • Humans
  • Liver Neoplasms / metabolism
  • Liver Neoplasms / pathology
  • Liver Neoplasms / virology
  • MicroRNAs / pharmacology*
  • Plasmids
  • RNA, Messenger / metabolism
  • Transfection
  • Virus Replication / drug effects*


  • DNA, Viral
  • Hepatitis B Surface Antigens
  • Hepatitis B e Antigens
  • MicroRNAs
  • RNA, Messenger