Suppression of CYP2C9 by microRNA hsa-miR-128-3p in human liver cells and association with hepatocellular carcinoma

Sci Rep. 2015 Feb 23;5:8534. doi: 10.1038/srep08534.

Abstract

Published studies have identified genetic variants, somatic mutations, and changes in gene expression profiles that are associated with hepatocellular carcinoma (HCC), particularly involving genes that encode drug metabolizing enzymes (DMEs). CYP2C9, one of the most abundant and important DMEs, is involved in the metabolism of many carcinogens and drugs and is down-regulated in HCC. To investigate the molecular mechanisms that control CYP2C9 expression, we applied integrative approaches including in silico, in vitro, and in vivo analyses to elucidate the role of microRNA hsa-miR-128-3p in the regulation of CYP2C9 expression and translation. RNA electrophoresis mobility shift assays demonstrated a direct interaction between hsa-miR-128-3p and its cognate target, the CYP2C9 transcript. Furthermore, the expression of a luciferase reporter gene containing the 3'-UTR of CYP2C9 and the endogenous expression of CYP2C9 were suppressed by transfection of hsa-miR-128-3p. Importantly, chemically-induced up- or down-regulation of hsa-miR-128-3p correlated inversely with the expression of CYP2C9. Finally, an association analysis revealed that the expression of hsa-miR-128-3p is inversely correlated with the expression of CYP2C9 in HCC tumor tissues. Altogether, the study helped to elucidate the mechanism of CYP2C9 regulation by hsa-miR-128-3p, and the inverse association in HCC.

MeSH terms

  • 3' Untranslated Regions
  • Base Sequence
  • Carcinoma, Hepatocellular / genetics*
  • Carcinoma, Hepatocellular / pathology
  • Cytochrome P-450 CYP2C9 / chemistry
  • Cytochrome P-450 CYP2C9 / genetics
  • Cytochrome P-450 CYP2C9 / metabolism*
  • Down-Regulation / drug effects
  • Electrophoretic Mobility Shift Assay
  • HEK293 Cells
  • Hep G2 Cells
  • Humans
  • Liver / metabolism*
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / pathology
  • MicroRNAs / metabolism*
  • RNA, Messenger / analysis
  • RNA, Messenger / metabolism
  • Sequence Alignment
  • Up-Regulation / drug effects
  • Zalcitabine / pharmacology

Substances

  • 3' Untranslated Regions
  • MIRN128 microRNA, human
  • MIRN143 microRNA, human
  • MicroRNAs
  • RNA, Messenger
  • Zalcitabine
  • Cytochrome P-450 CYP2C9