title: Proto-oncogenic isoform A2 of eukaryotic translation elongation factor eEF1 is a target of miR-663 and miR-744.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: State Key Laboratory on Molecular and Cell biology, Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kiev 03680, Ukraine.
authors.forename: A
authors.initials: A
authors.lastname: Vislovukh
authors.forename: G
authors.initials: G
authors.lastname: Kratassiouk
authors.forename: E
authors.initials: E
authors.lastname: Porto
authors.forename: N
authors.initials: N
authors.lastname: Gralievska
authors.forename: C
authors.initials: C
authors.lastname: Beldiman
authors.forename: G
authors.initials: G
authors.lastname: Pinna
authors.forename: A
authors.initials: A
authors.lastname: El'skaya
authors.forename: A
authors.initials: A
authors.lastname: Harel-Bellan
authors.forename: B
authors.initials: B
authors.lastname: Negrutskii
authors.forename: I
authors.initials: I
authors.lastname: Groisman
mesh_terms: D001943:Breast Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D048708:Cell Growth Processes / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D015536:Down-Regulation; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D035683:MicroRNAs / Q000008:administration & dosage* / Q000235:genetics; D020648:Peptide Elongation Factor 1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology; D014162:Transfection
chemical_list: C503234:EEF1A2 protein, human; C000715470:MAS1 protein, human; C552243:MIRN663 microRNA, human; C569733:MIRN744 microRNA, human; D035683:MicroRNAs; D020648:Peptide Elongation Factor 1; D000090063:Proto-Oncogene Mas; D012333:RNA, Messenger; D013267:Stilbenes; D000077185:Resveratrol