miRNA-340 inhibits osteoclast differentiation via repression of MITF

Abstract

Many miRNAs play critical roles in modulating various biological processes of osteoclast differentiation and function. Microphthalmia-associated transcription factor (MITF), a target of miR-340, served as pivotal transcription factor involved in osteoclast differentiation. However, the role of miR-340 and MITF during osteoclast differentiation has not yet been clearly established. Tartrate-resistant acid phosphatase (TRAP) staining assay was performed to identify osteoclasts differentiated from bone marrow-derived macrophages (BMMs). Quantitative reverse transcription PCR (qRT-PCR) or Western blotting was undertaken to examine the mRNA or protein expression respectively. Luciferase reporter assay was performed to investigate the interaction between miR-340 and MITF. MITF was knocked down and miR-340 was overexpressed and transfected into BMMs to detect their effects on osteoclast differentiation. Firstly, qRT-PCR analysis showed that miR-340 was down-regulated during osteoclast differentiation stimulated by macrophage-colony stimulating factor (M-CSF) and receptor activator of nuclear factor (NF)-κB (RANK) ligand (RANKL). Besides, we found that overexpression of miRNA-340 inhibited osteoclast differentiation and suppressed both the mRNA and protein level of MITF. Finally, Western blot and qRT-PCR analysis revealed that silencing MITF inhibited TRAP, calcitonin receptor, V-ATPase d2, and cathepsin K. miR-340 suppresses osteoclast differentiation by inhibiting MITF. Our findings may provide promising therapeutic targets for osteoclast-associated diseases.

Breast cancer cell‐derived exosomal miR‐20a‐5p promotes the proliferation and differentiation of osteoclasts by targeting SRCIN1

“…Recent studies have shown that miRNAs exert considerable roles in regulating various biological processes of osteoblast and osteoclast differentiation and function, thus making miRNAs as biomarkers and potential targets for osteoporosis therapy . Generally, miRNAs with upregulated expression level during osteoclast differentiation or formation tend to promote osteoclastogenesis, while miRNAs with downregulated level during osteoclast differentiation tend to inhibit osteoclastogenesis . A previous study using micro array analysis revealed the expression profile during different stages of murine osteoclastogenesis.…”

“…23 Generally, miRNAs with upregulated expression level during osteoclast differentiation or formation tend to promote osteoclastogenesis, while miRNAs with downregulated level during osteoclast differentiation tend to inhibit osteoclastogenesis. 24 A previous study using micro array analysis revealed the expression profile during different stages of murine osteoclastogenesis. Among all detected miRNAs, 49 were upregulated and 44 were downregulated, 25 however, miR-20a-5p was not among the list.…”

MITF—the first 25 years

“…In addition to transcriptional regulation, as might be expected for a key transcription factor, MITF is also subject to posttranscriptional regulation via both control of its mRNA polyadenylation by CPEB4 (Pérez-Guijarro et al 2016) and the action of microRNAs that play a key role in melanoma biology (Bell and Levy 2011;Kunz 2013). To date, several microRNAs have been described as inhibiting MITF expression, including miR-26a in melanoma (Qian et al 2017), miR-340 in osteoclasts (Zhao et al 2017), and miR-137 (Bemis et al 2008), miR-148 (Haflidadóttir et al 2010, miR-155 (Arts et al 2015), and miR-182 (Yan et al 2012). No doubt future research will identify additional miRs that can affect MITF expression and for now we refer the reader to the microRNA target prediction Web page for the 3 ′ untranslated region (UTR) (TargetScanHuman, http://www.targetscan.org/cgi-bin/ targetscan/vert_72/view_gene.cgi?rs=ENST00000328528 .6&taxid=9606&showcnc=0&shownc=0&shownc_nc=& showncf1=&showncf2=&subset=1).…”

CD226 deficiency on regulatory T cells aggravates renal fibrosis via up-regulation of Th2 cytokines through miR-340

“…found that miR‐340 inhibited endometrial carcinoma cell viability and proliferation by inducing apoptosis . Additionally, miR‐340 is involved in osteoclast differentiation by repressing microphthalmia‐associated transcription factor . Long noncoding RNA host gene 16 promoted osteosarcoma cell migration and invasion by sponging miRNA‐340 .…”

“…45 Additionally, miR-340 is involved in osteoclast differentiation by repressing microphthalmia-associated transcription factor. 46 Long noncoding RNA host gene 16 promoted osteosarcoma cell migration and invasion by sponging miRNA-340. 47 In addition, miR-340 was elevated in the maternal whole blood of patients with gestational diabetes and inversely regulated by insulin and glucose.…”