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Int. J. Mol. Sci. 2016, 17(2), 240; doi:10.3390/ijms17020240

Role of miR-222-3p in c-Src-Mediated Regulation of Osteoclastogenesis

1
Department of Biomedical Engineering, Indiana University—Purdue University Indianapolis, Indianapolis, IN 46202, USA
2
Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, Mie 514-8507, Japan
3
Department of Pharmacology, School of Dentistry, Aichi-Gakuin University, Nagoya 464-8650, Japan
*
Author to whom correspondence should be addressed.
Academic Editors: Y-h. Taguchi and Charles J. Malemud
Received: 15 December 2015 / Revised: 6 February 2016 / Accepted: 6 February 2016 / Published: 16 February 2016
(This article belongs to the Special Issue MicroRNA Regulation)
View Full-Text   |   Download PDF [3591 KB, uploaded 16 February 2016]   |  

Abstract

MicroRNAs (miRNAs) are small non-coding RNAs that play a mostly post-transcriptional regulatory role in gene expression. Using RAW264.7 pre-osteoclast cells and genome-wide expression analysis, we identified a set of miRNAs that are involved in osteoclastogenesis. Based on in silico analysis, we specifically focused on miR-222-3p and evaluated its role in osteoclastogenesis. The results show that the inhibitor of miR-222-3p upregulated the mRNA levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1) and tartrate-resistant acid phosphatase (TRAP), while its mimicking agent downregulated their mRNA levels. Western blot analysis showed that its inhibitor increased the protein levels of TRAP and cathepsin K, while its mimicking agent decreased their levels. Genome-wide mRNA expression analysis in the presence and absence of receptor activator of nuclear factor κ-B ligand (RANKL) predicted c-Src as a potential regulatory target of miR-222-3p. Live cell imaging using a fluorescence resonance energy transfer (FRET) technique revealed that miR-222-3p acted as an inhibitor of c-Src activity, and a partial silencing of c-Src suppressed RANKL-induced expression of TRAP and cathepsin K, as well as the number of multi-nucleated osteoclasts and their pit formation. Collectively, the study herein demonstrates that miR-222-3p serves as an inhibitor of osteoclastogenesis and c-Src mediates its inhibition of cathepsin K and TRAP. View Full-Text
Keywords: microarray; miRNA; osteoclastogenesis; RAW264.7 cells; c-Src microarray; miRNA; osteoclastogenesis; RAW264.7 cells; c-Src
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Takigawa, S.; Chen, A.; Wan, Q.; Na, S.; Sudo, A.; Yokota, H.; Hamamura, K. Role of miR-222-3p in c-Src-Mediated Regulation of Osteoclastogenesis. Int. J. Mol. Sci. 2016, 17, 240.

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