title: microRNA-495 inhibits chondrogenic differentiation in human mesenchymal stem cells by targeting Sox9.
abstract: The chondrogenic differentiation process of human mesenchymal stem cells (hMSCs) passes through multiple stages, which are carried out by various factors and their interactions. Recently, microRNAs that regulate chondrogenic differentiation have been reported. However, microRNA that regulates SRY-related high mobility group-box gene 9 (Sox9), a chondrogenic key factor, has not been identified in hMSC. In this study, we identified that microRNA-495 (miR-495) is an important regulator of hMSC chondrogenic differentiation. In our microarray, miR-495 was downregulated during transforming growth factor (TGF)-β3-induced chondrogenic differentiation of hMSCs in vitro. We found that there is an miR-495 binding site in the 3' untranslated region (3'UTR) of Sox9. We confirmed opposite expression between miR-495 and Sox9 by using real-time polymerase chain reaction. Further, overexpression of miR-495 inhibited Sox9 expression, and repression of miR-495 increased expression of Sox9 in SW1353 cells and hMSCs. Additionally, luciferase analysis revealed that miR-495 directly binds to the Sox9 3'UTR, and we confirmed a seed sequence of miR-495 on the Sox9 3'UTR. Subsequently, overexpression of miR-495 repressed the expression of the extracellular matrix (ECM) protein, such as type II collagen (Col2A1), aggrecan, and proteoglycan products, whereas inhibition of miR-495 increased their expression. Collectively, this study indicates that miR-495 directly targets Sox9, ultimately leading to the regulation of chondrogenic differentiation in hMSCs.
authors.affiliation: 1 Department of Orthopaedic Surgery, Yonsei University College of Medicine , Seoul, South Korea .
authors.forename: Seulgi
authors.initials: S
authors.lastname: Lee
authors.forename: Dong Suk
authors.initials: DS
authors.lastname: Yoon
authors.forename: Seungil
authors.initials: S
authors.lastname: Paik
authors.forename: Kyoung-Mi
authors.initials: KM
authors.lastname: Lee
authors.forename: Yeonsue
authors.initials: Y
authors.lastname: Jang
authors.forename: Jin Woo
authors.initials: JW
authors.lastname: Lee
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000328:Adult; D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D045744:Cell Line, Tumor; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics; D011509:Proteoglycans / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055758:SOX9 Transcription Factor / Q000235:genetics* / Q000378:metabolism; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology; D055815:Young Adult
chemical_list: D020413:3' Untranslated Regions; D015415:Biomarkers; D006025:Glycosaminoglycans; C577768:MIRN495 microRNA, human; D035683:MicroRNAs; D011509:Proteoglycans; D012333:RNA, Messenger; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D053782:Transforming Growth Factor beta3