title: MicroRNA-103a-3p controls proliferation and osteogenic differentiation of human adipose tissue-derived stromal cells.
abstract: The elucidation of the molecular mechanisms underlying the differentiation and proliferation of human adipose tissue-derived stromal cells (hADSCs) represents a critical step in the development of hADSCs-based cellular therapies. To examine the role of the microRNA-103a-3p (miR-103a-3p) in hADSCs functions, miR-103a-3p mimics were transfected into hADSCs in order to overexpress miR-103a-3p. Osteogenic differentiation was induced for 14 days in an osetogenic differentiation medium and assessed by using an Alizarin Red S stain. The regulation of the expression of CDK6 (cyclin-dependent kinase 6), a predicted target of miR-103a-3p, was determined by western blot, real-time PCR and luciferase reporter assays. Overexpression of miR-103a-3p inhibited the proliferation and osteogenic differentiation of hADSCs. In addition, it downregulated protein and mRNA levels of predicted target of miR-103a-3p (CDK6 and DICER1). In contrast, inhibition of miR-103a-3p with 2'O methyl antisense RNA increased the proliferation and osteogenic differentiation of hADSCs. The luciferase reporter activity of the construct containing the miR-103a-3p target site within the CDK6 and DICER1 3'-untranslated regions was lower in miR-103a-3p-transfected hADSCs than in control miRNA-transfected hADSCs. RNA interference-mediated downregulation of CDK6 and DICER1 in hADSCs inhibited their proliferation and osteogenic differentiation. The results of the current study indicate that miR-103a-3p regulates the osteogenic differentiation of hADSCs and proliferation of hADSCs by direct targeting of CDK6 and DICER1 partly. These findings further elucidate the molecular mechanisms governing the differentiation and proliferation of hADSCs.
authors.affiliation: Department of Physiology, School of Medicine, Pusan National University, Yangsan, Korea.
authors.forename: Da Sol
authors.initials: DS
authors.lastname: Kim
authors.affiliation: Department of Physiology, School of Medicine, Pusan National University, Yangsan, Korea.
authors.forename: Sun Young
authors.initials: SY
authors.lastname: Lee
authors.affiliation: Department of Physiology, School of Medicine, Pusan National University, Yangsan, Korea.
authors.forename: Jung Hee
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Plastic Surgery, School of Medicine, Pusan National University, Pusan, Korea.
authors.forename: Yong Chan
authors.initials: YC
authors.lastname: Bae
authors.affiliation: Department of Physiology, School of Medicine, Pusan National University, Yangsan, Korea.
authors.forename: Jin Sup
authors.initials: JS
authors.lastname: Jung
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D051361:Cyclin-Dependent Kinase 6 / Q000235:genetics; D053487:DEAD-box RNA Helicases / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D010012:Osteogenesis; D043244:Ribonuclease III / Q000235:genetics; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism
chemical_list: C544749:MIRN103 microRNA, human; D035683:MicroRNAs; D051361:Cyclin-Dependent Kinase 6; C408087:DICER1 protein, human; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases