miR-193b promotes cell proliferation by targeting Smad3 in human glioma

J Neurosci Res. 2014 May;92(5):619-26. doi: 10.1002/jnr.23339. Epub 2014 Feb 5.

Abstract

Studies have shown that several miRNAs play important roles in regulating a variety of cellular processes in gliomas. In these reports, upregulation of miR-193b has been found to be associated with a poor prognosis for glioma, but its functional mechanism in glioma remains unclear. This study investigates the roles of miR-193b in glioma tumor growth. We first showed that the expression of miR-193b was elevated in both glioma samples and glioma cells. Furthermore, downregulation of miR-193b by inhibitors was statistically correlated with a decrease in cell growth and a restored G1 accumulation. Luciferase assay and Western blot analysis revealed that Smad3 is a direct target of miR-193b. To prove that miR-193b regulated cell growth through the transforming growth factor-β (TGF-β) pathway in glioma cells by regulating Smad3, we tested endogenous targets of the TGF-β pathway by measuring the accumulation of p21 mRNAs after downregulation of miR-193b. The results confirmed that induction of p21 was promoted by miR-193b inhibitors in glioma cells, although this induction disappeared when Smad3 was knocked down with siRNA. Moreover, downregulation of Smad3 mitigates the miR-193b suppression of glioma proliferation. In conclusion, these results suggest that miR-193b regulated cell growth in glioma through the TGF-β pathway by regulating Smad3. Thus, our study indicates that miR-193b promotes cell proliferation by targeting Smad3 in human glioma, which may serve as a potentially useful target for development of miRNA-based therapies in the future.

Keywords: Smad3; cell proliferation; glioma; miR-193b; p21.

MeSH terms

  • Adult
  • Brain Neoplasms / pathology*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Female
  • Gene Expression Regulation, Neoplastic / drug effects
  • Gene Expression Regulation, Neoplastic / physiology*
  • Glioma / pathology*
  • HEK293 Cells
  • Humans
  • Male
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Oligonucleotides / genetics
  • Oligonucleotides / pharmacology
  • RNA, Small Interfering / metabolism
  • RNA, Small Interfering / pharmacology
  • Sincalide / pharmacology
  • Smad3 Protein / genetics
  • Smad3 Protein / metabolism*
  • Transfection
  • Transforming Growth Factor beta / metabolism
  • Tumor Cells, Cultured
  • Young Adult

Substances

  • MIRN193 microRNA, human
  • MicroRNAs
  • Oligonucleotides
  • RNA, Small Interfering
  • Smad3 Protein
  • Transforming Growth Factor beta
  • Sincalide