title: MicroRNA-194 reciprocally stimulates osteogenesis and inhibits adipogenesis via regulating COUP-TFII expression.
abstract: Osteoblasts and adipocytes are differentiated from common mesenchymal stem cells (MSCs) in processes which are tightly controlled by various growth factors, signaling molecules, transcriptional factors and microRNAs. Recently, chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) was identified as a critical regulator of MSC fate. In the present study, we aimed to identify some microRNAs (miR), which target COUP-TFII, and to determine the effects on MSCs fate. During osteoblastic or adipocytic differentiation from MSCs lineage cells, miR-194 expression was found to be reversal. In the cultures of mesenchymal C3H10T1/2 and primary bone marrow stromal cells, osteogenic stimuli increased miR-194 expression with accompanying decreases in COUP-TFII expression, whereas adipogenic stimuli reduced miR-194 expression with accompanying increases in COUP-TFII expression. A luciferase assay with COUP-TFII 3'-untranslated region (UTR) reporter plasmid, including the miR-194 binding sequences, showed that the introduction of miR-194 reduced the luciferase activity. However, it did not affect the activity of mutated COUP-TFII 3'-UTR reporter. Enforced expression of miR-194 significantly enhanced osteoblast differentiation, but inhibited adipocyte differentiation by decreasing COUP-TFII mRNA and protein levels. In contrast, inhibition of the endogenous miR-194 reduced matrix mineralization in the MSCs cultures, promoting the formation of lipid droplets by rescuing COUP-TFII expression. Furthermore, overexpression of COUP-TFII reversed the effects of miR-194 on the cell fates. Taken together, our results showed that miR-194 acts as a critical regulator of COUP-TFII, and can determinate the fate of MSCs to differentiate into osteoblasts and adipocytes. This suggests that miR-194 and COUP-TFII may be good target molecules for controlling bone and metabolic diseases.
authors.affiliation: 1] Research Center for Biomineralization Disorders, and Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, Korea [2] Department of Pharmacology and Dental Therapeutics, School of Dentistry, Chonnam National University, Gwangju, Korea.
authors.forename: B-C
authors.initials: BC
authors.lastname: Jeong
authors.affiliation: 1] Research Center for Biomineralization Disorders, and Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, Korea [2] Department of Pharmacology and Dental Therapeutics, School of Dentistry, Chonnam National University, Gwangju, Korea.
authors.forename: I-H
authors.initials: IH
authors.lastname: Kang
authors.affiliation: 1] Research Center for Biomineralization Disorders, and Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, Korea [2] Department of Conservative Dentistry, School of Dentistry, Chonnam National University, Gwangju, Korea.
authors.forename: Y-C
authors.initials: YC
authors.lastname: Hwang
authors.affiliation: 1] Research Center for Biomineralization Disorders, and Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, Korea [2] Department of Oral Anatomy, School of Dentistry, Chonnam National University, Gwangju, Korea.
authors.forename: S-H
authors.initials: SH
authors.lastname: Kim
authors.affiliation: 1] Research Center for Biomineralization Disorders, and Dental Science Research Institute, School of Dentistry, Chonnam National University, Gwangju, Korea [2] Department of Pharmacology and Dental Therapeutics, School of Dentistry, Chonnam National University, Gwangju, Korea.
authors.forename: J-T
authors.initials: JT
authors.lastname: Koh
mesh_terms: D020413:3' Untranslated Regions; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D050156:Adipogenesis / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D051837:COUP Transcription Factor II / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D003470:Culture Media / Q000494:pharmacology; D005269:Femur / Q000166:cytology; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics; D061251:Primary Cell Culture; D015398:Signal Transduction; D013977:Tibia / Q000166:cytology
chemical_list: D020413:3' Untranslated Regions; C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D051837:COUP Transcription Factor II; D003470:Culture Media; C554845:MIRN194 microRNA, mouse; D035683:MicroRNAs; C497587:Nr2f2 protein, mouse; D008156:Luciferases