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

International Journal of Molecular Sciences
Shinya TakigawaKazunori Hamamura

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-nucleate...Continue Reading

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Apr 11, 2017·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Dong-Yang QianHong Xia
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Datasets Mentioned

BETA
GSE74847

Methods Mentioned

BETA
FRET
biosensor
PCA
PCR
transfection
Fluorescence
biosensors

Software Mentioned

miRWalk
Dcstamp
Image J
Microt4
PITA
miRanda
miRMap
RNA22
RNAhybrid
Targetscan

Related Concepts

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