MicroRNA-590-5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation

M. Vishal, S. Vimalraj, R. Ajeetha, M. Gokulnath, R. Keerthana, Z. He, Nicola Partridge, N. Selvamurugan

Research output: Contribution to journalArticle

Abstract

Mesenchymal stem cells (MSCs) are multipotent cells and their differentiation into the osteoblastic lineage is strictly controlled by several regulators, including microRNAs (miRNAs). Runx2 is a bone transcription factor required for osteoblast differentiation. Here, we used in silico analysis to identify a number of miRNAs that putatively target Runx2 and its co-factors to mediate both positive and negative regulation of osteoblast differentiation. Among these miRNAs, miR-590-5p was selected and its expression was found to be increased during osteoblast differentiation. When mouse MSCs (mMSCs) were transiently transfected with a miR-590-5p mimic, we detected an increase in both calcium deposition and the mRNA expression of osteoblast differentiation marker genes such as alkaline phosphatase (ALP) and type I collagen genes. Smad7 was found to be among the putative target genes of miR-590-5p and its mRNA and protein expression decreased after miR-590-5p mimic transfection in human osteoblast-like cells (MG63). Our analysis indicated that Runx2 was not a putative target of miR-590-5p. However, Runx2 protein, but not mRNA expression, increased after miR-590-5p mimic transfection in MG63 cells. Runx2 protein expression was increased with knockdown of Smad7 expression by Smad7 siRNA in these cells. We further identified that the 3′-untranslated region of Smad7 was directly targeted by miR-590-5p; this was done using the luciferase reporter gene system. It is known that Smad7 inhibits osteoblast differentiation via Smurf2-mediated Runx2 degradation. Hence, based on our results, we suggest that miR-590-5p promotes osteoblast differentiation by indirectly protecting and stabilizing the Runx2 protein by targeting Smad7 gene expression. J. Cell. Physiol. 232: 371–380, 2017.

Original languageEnglish (US)
Pages (from-to)371-380
Number of pages10
JournalJournal of Cellular Physiology
Volume232
Issue number2
DOIs
StatePublished - Feb 1 2017

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Osteoblasts
MicroRNAs
Core Binding Factor Alpha 1 Subunit
Genes
Messenger RNA
Transfection
Differentiation Antigens
3' Untranslated Regions
Protein Transport
Collagen Type I
Stem cells
Luciferases
Mesenchymal Stromal Cells
Reporter Genes
Gene expression
Computer Simulation
Small Interfering RNA
Alkaline Phosphatase
Cell Differentiation
Bone

ASJC Scopus subject areas

  • Medicine(all)
  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Vishal, M., Vimalraj, S., Ajeetha, R., Gokulnath, M., Keerthana, R., He, Z., ... Selvamurugan, N. (2017). MicroRNA-590-5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation. Journal of Cellular Physiology, 232(2), 371-380. https://doi.org/10.1002/jcp.25434

MicroRNA-590-5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation. / Vishal, M.; Vimalraj, S.; Ajeetha, R.; Gokulnath, M.; Keerthana, R.; He, Z.; Partridge, Nicola; Selvamurugan, N.

In: Journal of Cellular Physiology, Vol. 232, No. 2, 01.02.2017, p. 371-380.

Research output: Contribution to journalArticle

Vishal, M, Vimalraj, S, Ajeetha, R, Gokulnath, M, Keerthana, R, He, Z, Partridge, N & Selvamurugan, N 2017, 'MicroRNA-590-5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation', Journal of Cellular Physiology, vol. 232, no. 2, pp. 371-380. https://doi.org/10.1002/jcp.25434
Vishal M, Vimalraj S, Ajeetha R, Gokulnath M, Keerthana R, He Z et al. MicroRNA-590-5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation. Journal of Cellular Physiology. 2017 Feb 1;232(2):371-380. https://doi.org/10.1002/jcp.25434
Vishal, M. ; Vimalraj, S. ; Ajeetha, R. ; Gokulnath, M. ; Keerthana, R. ; He, Z. ; Partridge, Nicola ; Selvamurugan, N. / MicroRNA-590-5p Stabilizes Runx2 by Targeting Smad7 During Osteoblast Differentiation. In: Journal of Cellular Physiology. 2017 ; Vol. 232, No. 2. pp. 371-380.
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