MicroRNA-223 Suppresses Osteoblast Differentiation by Inhibiting DHRS3

被引：39

作者：

Zhang, Shijie ^{[1
,2
,3
]}

Liu, Yi ^{[2
,3
]}

Zheng, Zhong ^{[4
,5
,6
]}

Zeng, Xuemin ^{[2
,3
]}

Liu, Dongxu ^{[2
,3
]}

Wang, Chunling ^{[2
,3
]}

Ting, Kang ^{[4
,5
,6
]}

机构：

[1] Shandong Univ, Qilu Hosp, Dept Orthodont, Jinan, Shandong, Peoples R China

[2] Shandong Univ, Sch Stomatol, Shandong Prov Key Lab Oral Tissue Regenerat, Jinan 250012, Shandong, Peoples R China

[3] Shandong Univ, Sch Stomatol, Dept Orthodont, Jinan 250012, Shandong, Peoples R China

[4] Univ Calif Los Angeles, Sch Dent, Sect Orthodont, Div Growth & Dev, Los Angeles, CA 90024 USA

[5] Univ Calif Los Angeles, Orthopaed Hosp Res Ctr, Div Plast & Reconstruct Surg, Los Angeles, CA USA

[6] Univ Calif Los Angeles, Dept Orthopaed Surg, Orthopaed Hosp Res Ctr, Los Angeles, CA USA

来源：

CELLULAR PHYSIOLOGY AND BIOCHEMISTRY | 2018年 / 47卷 / 02期

基金：

中国国家自然科学基金; 中国博士后科学基金;

关键词：

miR-223; DHRS3; BMSC; Osteogenic differentiation; MESENCHYMAL STEM-CELLS; BONE MORPHOGENETIC PROTEIN; RETINOIC ACID; OSTEOGENIC DIFFERENTIATION; MARROW; EXPRESSION; PROLIFERATION; OSTEOSARCOMA; METABOLISM; ARTHRITIS;

D O I：

10.1159/000490021

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

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. (C) 2018 The Author(s) Published by S. Karger AG, Basel

引用

页码：667 / 679

页数：13

共 32 条

[1] The Retinaldehyde Reductase Activity of DHRS3 Is Reciprocally Activated by Retinol Dehydrogenase 10 to Control Retinoid Homeostasis [J].