Research Article

 Blocking the expression of the hepatitis B virus S gene in hepatocellular carcinoma cell lines with an anti-gene locked nucleic acid in vitro

Published: May 22, 2015
Genet. Mol. Res. 14 (2) : 5445-5451 DOI: https://doi.org/10.4238/2015.May.22.14
Cite this Article:
(2015).  Blocking the expression of the hepatitis B virus S gene in hepatocellular carcinoma cell lines with an anti-gene locked nucleic acid in vitro. Genet. Mol. Res. 14(2): gmr5483. https://doi.org/10.4238/2015.May.22.14
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Abstract

The aim of this study was to investigate the effects of inhibition of the hepatitis B virus (HBV) S gene by polypurine region locked nucleic acid on viral replication in cells. We designed and synthe­sized a locked nucleic acid, phosphorothioate oligonucleotides, unmod­ified oligonucleotides, and unrelated control sequence for the hepatitis B virus S gene polypurine region. HepG2.2.15 cells were transfected by cationic liposome, and fluorescence quantitative polymerase chain reaction technology (PCR) and time-resolved fluoroimmunoassay tech­nology was utilized to monitor the content of HBV DNA, HbsAg, and HBeAg at 2, 4, 6, 8 and 10 days post-transfection. The effects on cell metabolism were detected by four methyl thiazolyl tetrazolium assay. The locked nucleic acid had an obvious effect on HBV DNA replication and HBsAg and HBeAg expression in a dose and time dependent man­ner. The inhibition rates were 52.14, 57.48, and 29.63% after 6 days, respectively. The locked nucleic acid had no significant effect on cell metabolism. The HBV S gene polypurine region locked nucleic acid could effectively inhibit the replication of HBV in vitro, and could pro­vide an effective target for the treatment of HBV and a theoretical and experimental basis for anti-gene therapy.

The aim of this study was to investigate the effects of inhibition of the hepatitis B virus (HBV) S gene by polypurine region locked nucleic acid on viral replication in cells. We designed and synthe­sized a locked nucleic acid, phosphorothioate oligonucleotides, unmod­ified oligonucleotides, and unrelated control sequence for the hepatitis B virus S gene polypurine region. HepG2.2.15 cells were transfected by cationic liposome, and fluorescence quantitative polymerase chain reaction technology (PCR) and time-resolved fluoroimmunoassay tech­nology was utilized to monitor the content of HBV DNA, HbsAg, and HBeAg at 2, 4, 6, 8 and 10 days post-transfection. The effects on cell metabolism were detected by four methyl thiazolyl tetrazolium assay. The locked nucleic acid had an obvious effect on HBV DNA replication and HBsAg and HBeAg expression in a dose and time dependent man­ner. The inhibition rates were 52.14, 57.48, and 29.63% after 6 days, respectively. The locked nucleic acid had no significant effect on cell metabolism. The HBV S gene polypurine region locked nucleic acid could effectively inhibit the replication of HBV in vitro, and could pro­vide an effective target for the treatment of HBV and a theoretical and experimental basis for anti-gene therapy.