In vitro inhibition of hepatitis C virus by antisense oligonucleotides in PBMC compared to hepatoma cells

Biomed Res Int. 2014;2014:196712. doi: 10.1155/2014/196712. Epub 2014 Jun 1.

Abstract

Aim: To assess the efficiency of phosphorothioate antisense oligodeoxynucleotide 1 (S-ODN1) on HCV translation inhibition in PBMC compared to hepatoma cells in vitro for the first time.

Materials and methods: The study included 34 treatment naive HCV patients. IRES domain III and IV sequence variations were tested in 45 clones from 9 HCV patients. PBMC of HCV positive patients were subjected to S-ODN in vitro. Concomitantly HepG2 cells infected by the same patient's serum were also treated with S-ODN1 for 24 and 48 hours. Cellular RNA was tested for HCV plus and minus strands by reverse transcription polymerase chain reaction (RT-PCR).

Results: Sequence variations were seen in HCV IRES domain III only while domain IV was conserved among all the tested patient's clones. S-ODN1 successfully inhibited HCV translation in HepG2 cells, while in PBMC inhibition was partial.

Conclusion: HCV IRES domain IV is more conserved than domain IIId in genotype 4 HCV patients. S-ODN against HCV IRES domain IV was not efficient to inhibit HCV translation in PBMC under the study conditions. Further studies testing other S-ODN targeting other HCV IRES domains in PBMC should be done.

MeSH terms

  • 5' Untranslated Regions / genetics
  • Carcinoma, Hepatocellular / genetics*
  • Carcinoma, Hepatocellular / pathology
  • Carcinoma, Hepatocellular / therapy
  • Carcinoma, Hepatocellular / virology
  • Hep G2 Cells
  • Hepacivirus / drug effects
  • Hepacivirus / genetics*
  • Humans
  • Leukocytes, Mononuclear / virology
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / pathology
  • Liver Neoplasms / therapy
  • Liver Neoplasms / virology
  • Oligonucleotides, Antisense / administration & dosage
  • Oligonucleotides, Antisense / genetics*
  • Protein Biosynthesis
  • RNA, Viral / antagonists & inhibitors
  • RNA, Viral / genetics
  • Virus Internalization

Substances

  • 5' Untranslated Regions
  • Oligonucleotides, Antisense
  • RNA, Viral