Proto-oncogenic isoform A2 of eukaryotic translation elongation factor eEF1 is a target of miR-663 and miR-744

Br J Cancer. 2013 Jun 11;108(11):2304-11. doi: 10.1038/bjc.2013.243. Epub 2013 May 21.

Abstract

Background: Eukaryotic translation elongation factor 1A2 (eEF1A2) is a known proto-oncogene. We proposed that stimulation of the eEF1A2 expression in cancer tissues is caused by the loss of miRNA-mediated control.

Methods: Impact of miRNAs on eEF1A2 at the mRNA and protein levels was examined by qPCR and western blot, respectively. Dual-luciferase assay was applied to examine the influence of miRNAs on 3'-UTR of EEF1A2. To detect miRNA-binding sites, mutations into the 3'-UTR of EEF1A2 mRNA were introduced by the overlap extension PCR.

Results: miR-663 and miR-744 inhibited the expression of luciferase gene attached to the 3'-UTR of EEF1A2 up to 20% and 50%, respectively. In MCF7 cells, overexpression of miR-663 and miR-744 reduced the EEF1A2 mRNA level by 30% and 50%. Analogous effects were also observed at the eEF1A2 protein level. In resveratrol-treated MCF7 cells the upregulation of mir-663 and mir-744 was accompanied by downregulation of EEF1A2 mRNA. Both miRNAs were able to inhibit the proliferation of MCF7 cells.

Conclusion: miR-663 and miR-744 mediate inhibition of the proto-oncogene eEF1A2 expression that results in retardation of the MCF7 cancer cells proliferation. Antitumour effect of resveratrol may include stimulation of the miR-663 and miR-744 expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / enzymology*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / pathology
  • Breast Neoplasms / therapy*
  • Cell Growth Processes / genetics
  • Cell Movement / genetics
  • Down-Regulation
  • Female
  • Humans
  • MCF-7 Cells
  • MicroRNAs / administration & dosage*
  • MicroRNAs / genetics
  • Peptide Elongation Factor 1 / antagonists & inhibitors
  • Peptide Elongation Factor 1 / biosynthesis
  • Peptide Elongation Factor 1 / genetics*
  • Proto-Oncogene Mas
  • RNA, Messenger / antagonists & inhibitors
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Resveratrol
  • Stilbenes / pharmacology
  • Transfection

Substances

  • EEF1A2 protein, human
  • MAS1 protein, human
  • MIRN663 microRNA, human
  • MIRN744 microRNA, human
  • MicroRNAs
  • Peptide Elongation Factor 1
  • Proto-Oncogene Mas
  • RNA, Messenger
  • Stilbenes
  • Resveratrol