Abstract

To study the effects of microRNA‐98 (miR‐98) on human bone mesenchymal stromal cells (hBMSCs). The patients undergoing hip arthroplasty were selected by inclusion/exclusion criteria for this study. The extracted hBMSCs were detected of osteogenic differentiation by alizarin red S staining, and of cell phenotype by flow cytometry. Bioinformatics, dual luciferase report, western blotting, RT‐PCR and immunoblotting were used in our study. The hBMSCs were divided into miR‐98 mimics, miR‐98 negative control (NC), miR‐98 inhibitors, Mock and miR‐98 inhibitors + siBMP2 groups. Human bone mesenchymal stromal cells were extracted and purified in vitro and had specific cytological morphology, surface markers and abilities of self‐renewal and differentiation. Compared with the NC group and Mock group, the miR‐98 mimics group showed increased miR‐98 level while the miR‐98 inhibitors group decreased miR‐98 level (both P < 0.01). Dual luciferase reporter showed BMP2 was the target gene of miR‐98. The levels of mRNA and protein expression of BMP2, protein expression of RUNX2, alkaline phosphatase activity and osteocalcin content significantly decreased in the miR‐98 mimics group while increased in the miR‐98 inhibitors group and showed no changes in the NC group and Mock group (all P < 0.05). The miR‐98 mimics group showed obviously declined stained red particles and the miR‐98 inhibitors group showed opposite result. After lowering the expression of miR‐98, osteogenic differentiation ability of hBMSCs rose, which was weakened by the transfection with siBMP2. miR‐98 may regulate osteogenic differentiation of hBMSCs by targeting BMP2.

Highlights

  • Human bone mesenchymal stromal cells are a group of clonogenic cells in the bone marrow stroma and is capable of self-renewal and multilineage differentiation into such cell types as mesoderm-type cells, including osteoblast, adipocyte and chondrocytes, and epithelial cells [1,2,3]

  • No direct research on miR-98 regulating osteogenic differentiation was found, its effect on cell growth may have links with skeletal cells; we carried out the present study based on the hypothesis that miR-98 may participate in the mechanism of osteogenic differentiation of human bone mesenchymal stromal cells (hBMSCs)

  • These results indicated that the inhibitory effect of miR-98 on the osteogenic differentiation of hBMSCs was mainly caused by its inhibition of BMP2 signalling pathway

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Summary

Introduction

Human bone mesenchymal stromal cells ( known as mesenchymal stem cells, bone marrow stromal cells or multipotent mesenchymal stromal cells) are a group of clonogenic cells in the bone marrow stroma and is capable of self-renewal and multilineage differentiation into such cell types as mesoderm-type cells, including osteoblast, adipocyte and chondrocytes, and epithelial cells [1,2,3]. According to Papaioannou et al, extracellular signalling molecules such as miRNA can trigger bone cells to proliferate and differentiate, which regulates gene expression and monitor cellular functions [4]. Several miRNAs were demonstrated to get involved in osteogenic differentiation modulation: miR-125b negatively regulated osteogenic differentiation by targeting ErbB2, VDR and Osterix; miR133 and miR-135 inhibited differentiation of mouse osteoprogenitors, respectively, by targeting RUNX2 and recognizing SMAD5; miR-26a and miR-29b promoted osteogenic differentiation of human adipose tissue-derived stem cells, and positively regulated mouse osteoblast differentiation [12,13,14,15,16]. Identified targets of let-7 include cell cycle regulators, promoters of growth and multiple early embryonic genes [23,24,25,26,27]. No direct research on miR-98 regulating osteogenic differentiation was found, its effect on cell growth may have links with skeletal cells; we carried out the present study based on the hypothesis that miR-98 may participate in the mechanism of osteogenic differentiation of hBMSCs

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