Igf2-derived intronic miR-483 promotes mouse hepatocellular carcinoma cell proliferation

Mol Cell Biochem. 2012 Feb;361(1-2):337-43. doi: 10.1007/s11010-011-1121-x. Epub 2011 Nov 4.

Abstract

Most intronic micro-RNAs are coexpressed with their host genes, suggesting that they may play similar roles. The function of miR-483 remains unknown, although it is embedded in an intron of Igf2 gene, which is an activator of hepatocellular carcinoma proliferation. In the present study, we provide evidence that Igf2-derived miR-483 can induce proliferation in hepatocellular carcinoma cells. The miR-483 promotion of proliferation was analysed by soft agar colony formation assay and proliferation curve assay. The effect of miR-483 on Socs3 expression was examined by Western blot and a reporter assay. Our results revealed that Igf2-derived intronic miR-483 was identified by the application of 94G6, an inhibitor of Igf2 at the transcriptional level. All results from the (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) MTT assay, the proliferation curve assay and the soft agar colony formation assay showed that miR-483 promoted the proliferation of hepatocellular carcinoma cells. Finally, Socs3, a putative target predicted by bioinformatics, was regulated by miR-483 at mRNA and protein levels. Direct binding with the 3' UTR was identified by a luciferase activity assay. Our findings demonstrate that Igf2-derived intronic miR-483, through downregulation of its target Socs3, regulates hepatoma cell proliferation and thus may serve as a potential target for hepatocellular carcinoma therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Base Sequence
  • Carcinoma, Hepatocellular
  • Cell Line, Tumor
  • Cell Proliferation*
  • Cell Survival
  • Gene Expression
  • Gene Expression Regulation, Neoplastic
  • Gene Silencing
  • Insulin-Like Growth Factor II / genetics*
  • Introns*
  • Liver Neoplasms
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Suppressor of Cytokine Signaling 3 Protein
  • Suppressor of Cytokine Signaling Proteins / genetics*
  • Suppressor of Cytokine Signaling Proteins / metabolism

Substances

  • 3' Untranslated Regions
  • IGF2 protein, mouse
  • MicroRNAs
  • Mirn483 microRNA, mouse
  • Socs3 protein, mouse
  • Suppressor of Cytokine Signaling 3 Protein
  • Suppressor of Cytokine Signaling Proteins
  • Insulin-Like Growth Factor II