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

  • Ali Asgharzadeh Department of Hematology, Imam Reza Hospital, Mashhad University of Medical Sciences, Mashhad, Iran
  • Shaban Alizadeh Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Reza Keramati Cancer Molecular Pathology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Masoud Soleimani Department of Hematology, Tarbiat Modares University, Tehran, Iran
  • Amir Atashi Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Shahroud University of Medical Sciences, Shahroud, Iran
  • Mahdi Edalati Department of Laboratory Sciences, Paramedical Faculty, Tabriz University of Medical Sciences, Tabriz, Iran Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
  • Zahra Kashani Khatib Hematology Department, High Institute for Research and Education in Transfusion Medicine, Iranian Blood Transfusion Organization (IBTO), Tehran, Iran
  • Mohammad Rafiee Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Mohyedin Barzegar Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Hengamehsadat Razavi Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
Keywords: microRNAs, miR-210, MSC, osteoblast, cord blood, cell differentiation

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.

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Author Biographies

Ali Asgharzadeh, Department of Hematology, Imam Reza Hospital, Mashhad University of Medical Sciences, Mashhad, Iran
Department of Hematology
Shaban Alizadeh, Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
Department of Hematology
Masoud Soleimani, Department of Hematology, Tarbiat Modares University, Tehran, Iran
Department of Hematology
Amir Atashi, Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Shahroud University of Medical Sciences, Shahroud, Iran
Department of Medical Laboratory Sciences
Mahdi Edalati, Department of Laboratory Sciences, Paramedical Faculty, Tabriz University of Medical Sciences, Tabriz, Iran Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran
Department of Laboratory Sciences
Zahra Kashani Khatib, Hematology Department, High Institute for Research and Education in Transfusion Medicine, Iranian Blood Transfusion Organization (IBTO), Tehran, Iran
Hematology Department
Mohammad Rafiee, Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Department of Hematology and Blood Banking
Mohyedin Barzegar, Department of Hematology and Blood Banking, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Department of Hematology and Blood Banking
Hengamehsadat Razavi, Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran
Department of Hematology

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Published
2018-11-07
How to Cite
1.
Asgharzadeh A, Alizadeh S, Keramati MR, Soleimani M, Atashi A, Edalati M, Kashani Khatib Z, Rafiee M, Barzegar M, Razavi H. Upregulation of miR-210 promotes differentiation of mesenchymal stem cells (MSCs) into osteoblasts. Bosn J of Basic Med Sci [Internet]. 2018Nov.7 [cited 2019Aug.26];18(4):328-35. Available from: https://bjbms.org/ojs/index.php/bjbms/article/view/2633
Section
Molecular Biology