Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts

Asgharzadeh, A. and Alizadeh, S. and Keramati, M. R. and Soleimani, M. and Atashi, A. and Edalati, M. and Khatib, Z. K. and Rafiee, M. and Barzegar, M. and Razavi, H. (2018) Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts. Bosnian Journal of Basic Medical Sciences, 18 (4). pp. 328-335.

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Abstract

Numerous studies indicated that microRNAs are critical in the regulation of cellular differentiation, by controlling the expression of underlying genes. The aim of this study was to investigate the effect of miR-210 upregulation on differentiation of human umbilical cord blood (HUCB)-derived mesenchymal stem cells (MSCs) into osteoblasts. MSCs were isolated from HUCB and confirmed by their adipogenic/osteogenic differentiation and flow cytometric analysis of surface markers. Pre-miR-210 was amplified from human DNA, digested and ligated with plenti-III-mir-green fluorescent protein (GFP) vector, and cloned in STBL4 bacteria. After confirmation with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), the plenti-III-GFP segment bearing pre-miR-210 was transfected into MSCs by electroporation. Two control vectors, pmaxGFP and Scramble, were transfected separately into MSCs. The expression of miR-210 and genes related to osteoblast differentiation, i.e. runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteocalcin gene, in the three groups of transfected MSCs was analyzed 0, 7, 14, and 21 days of transfection by quantitative reverse transcription PCR (qRT-PCR). Overexpression of miR-210 was observed in MSCs transfected with miR-210-bearing plasmid, and this was significantly different compared to Scramble group (p < 0.05). Significantly increased expression of Runx2 (at day 7 and 14), ALP and osteocalcin genes (at all time points for both genes) was observed in MSCs with miR-210-bearing plasmid compared to controls. Overall, the overexpression of miR-210 in MSCs led to MSC differentiation into osteoblasts, most probably by upregulating the Runx2, ALP, and osteocalcin genes at different stages of cell differentiation. Our study confirms the potential of miRNAs in developing novel therapeutic strategies that could target regulatory mechanisms of cellular differentiation in various disease states. © 2018 ABMSFBIH.

Item Type: Article
Additional Information: Cited By :3 Export Date: 16 February 2020 Correspondence Address: Keramati, M.R.; Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Avicenna Street, Iran; email: Keramatimr@mums.ac.ir
Uncontrolled Keywords: Cell differentiation Cord blood MicroRNAs MiR-210 MSC Osteoblast green fluorescent protein microRNA MIRN210 microRNA, human osteocalcin RUNX2 protein, human transcription factor RUNX2 biosynthesis bone development drug effect genetics human mesenchymal stem cell restriction fragment length polymorphism upregulation Core Binding Factor Alpha 1 Subunit Green Fluorescent Proteins Humans Mesenchymal Stem Cells Osteoblasts Osteogenesis Polymorphism, Restriction Fragment Length Up-Regulation
Subjects: QU Biochemistry
QZ pathology-neoplasms-Genetics
Divisions: Mashhad University of Medical Sciences
Depositing User: lib2 lib2 lib2
Date Deposited: 06 Jun 2020 08:18
Last Modified: 06 Jun 2020 08:18
URI: http://eprints.mums.ac.ir/id/eprint/17209

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