MicroRNA-223 Suppresses Osteoblast Differentiation by Inhibiting DHRS3

Abstract

Background/Aims: In this study, we aimed to use bioinformatics tools to identify the specific miRNAs and mRNAs involved in osteogenic differentiation and to further explore the way in which miRNA regulates osteogenic differentiation. Methods: The microarray GSE80614, which includes data from 3 human mesenchymal stromal cells (hMSCs) and 3 hMSCs after 72 hours (hr) of osteogenic differentiation, was used to screen for differentially expressed mRNAs. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses of these mRNAs were conducted using Gene Set Enrichment Analysis (GSEA). Then, the miRanda website was employed to detect the binding sites of DHRS3. In vitro experiments, including RT-PCR and western blotting, were used to detect miR-233 and DHRS3 expression levels 7 and 14 days (d) after the induction of osteogenic differentiation using human bone marrow-derived mesenchymal stem cells (hBMSCs). The target relationship between miR-223 and DHRS3 was confirmed by a dual luciferase assay. ALP (alkaline phosphatase) staining, ARS (Alizarin Red S) staining and western blotting (Runx2, OPN, OCN) were used to detect the level of osteogenic differentiation after transfection with miR-223 mimics and DHRS3 cDNA. Results: In this study, 127 mRNAs differentially expressed during osteogenic differentiation were identified in GSE80614. GO term and KEGG pathway enrichment analyses found that the retinol metabolism pathway was activated during osteogenic differentiation and that DHRS3, which is involved in the pathway, was upregulated. During osteogenic differentiation in hBMSCs, miR-223 was gradually downregulated, while DHRS3 was upregulated. After 14 days of osteogenic differentiation, ALP and ARS staining assay results showed strong ALP activity and extracellular matrix calcification with the inhibition of miR-223 or the overexpression of DHRS3. Furthermore, the expression levels of Runx2, OPN, and OCN were upregulated with the knockdown of miR-223 or the overexpression of DHRS3, while the simultaneous transfection of a miR-223 agomir and DHRS3 cDNA resulted in no significant difference from the negative control (NC) group. Conclusion: The inhibition of miR-223 promotes the osteogenic differentiation of hBMSCs via the upregulation of DHRS3.

MicroRNA‑338‑3p regulates age‑associated osteoporosis via targeting PCSK5

“…The findings revealed that miR-338-3p showed an opposite trend of expression in BMSCs with age and exerted negative functions on PCSK5, thereby limiting bone formation. Several studies also reported that an increasing number of miRNAs could regulate BMSC differentiation (31)(32)(33). In the present study, increased expression of miR-338-3p was observed in BMSCs of older rat samples, which indicated a counter-productive role of miR-338-3p in osteogenesis.…”

<p>Zirconium Oxide Thin Films Obtained by Atomic Layer Deposition Technology Abolish the Anti-Osteogenic Effect Resulting from miR-21 Inhibition in the Pre-Osteoblastic MC3T3 Cell Line</p>

“…MiR-223 was recently identified as a key modulator of osteoblastic differentiation in MC3T3-E1 cells, which targets histone deacetylase 2 (HDAC2). 55 To summarize, the obtained ZrO 2 ALD coatings are cytocompatible toward the MC3T3 osteoblastic cell line. The coatings promote the metabolic potential and may mitigate the effect of miR-21 inhibition, increasing expression of osteogenic markers, both at mRNA and miRNA level.…”

A Novel OsteomiRs Expression Signature for Osteoblast Differentiation of Human Amniotic Membrane-Derived Mesenchymal Stem Cells

“…Several miRNAs modulate osteogenic differentiation; thus they have been dubbed as osteomiRs. For instance, miR-125b regulates osteoblastic differentiation of human MSCs through targeting of BMPR1b [20], whereas miR-223 suppresses osteoblast differentiation by inhibiting DHRS3 [21]. However, miRNAs functions during differentiation of hAM-MSCs to osteogenic lineage have not been completely explored.…”