Influence of MicroRNA-141 on Inhibition of the Proliferation of Bone Marrow Mesenchymal Stem Cells in Steroid-Induced Osteonecrosis via SOX11.

Chen‐Yang Meng,Shi‐Bing Guo,Ting Hao,Wei Feng,Fei Xue,Zhen‐Qun Zhao

doi:10.1111/os.12603

Abstract

ObjectiveTo investigate whether miR‐141 and the sex determination region of Y chromosome box 11 (SOX11) play roles in steroid‐induced avascular necrosis of the femoral head (SANFH), and to explore whether miR‐141 could target SOX11 to influence the proliferation of bone marrow mesenchymal stem cells (BMSC).MethodsBone marrow mesenchymal stem cells (BMSC) were isolated and cultured from 4‐week‐old Sprague Dawley rats. A flow cytometry assay was performed to identify BMSC. BMSC were divided into two groups: a control group and a dexamethasone (DEX) group. BMSC were transfected by miR‐141 mimic, miR‐141 inhibitor, and SOX11. Real‐time polymerase chain reaction (PCR) assay was performed to investigate the mRNA expression of miR‐141 and SOX11. The results were used to determine the effect of transfection and to verify the expression in each group and the association between miR‐141 and SOX11. Luciferase reporter assay revealed the targeted binding site between miR‐141 and the 3′‐untranslated region of SOX11 mRNA. MTT assays were performed to investigate the proliferation of BMSC in the miR‐141 mimic, miR‐141 inhibitor, and SOX11 groups.ResultThe results of the flow cytometry assay suggested that cells were positive for CD29 and CD90 while negative for CD45. This meant that the isolated and cultured cells were not hematopoietic stem cells. In addition, cell transfection was successful based on the expression of miR‐141 and SOX11. According to the results of real‐time PCR assay, the mRNA expression of miR‐141 in SANFH was upregulated (4.117 ± 0.042 vs 1 ± 0.027, P < 0.001), while SOX11 was downregulated (0.611 ± 0.055 vs 1 ± 0.027, P < 0.001) compared with the control group. Based on the results of the luciferase experiment, MiR‐141 could directly target the expression of SOX11. Inhibition of miR‐141 could upregulate the expression of SOX11 (2.623 ± 0.220 vs 1 ± 0.095, P < 0.001) according to the results of a real‐time PCR assay. MiR‐141 inhibited the proliferation of BMSC (0.618 ± 0.092 vs 1.004 ± 0.082, P < 0.001), while suppression of miR‐141 increased the proliferation of BMSC (0.960 ± 0.095 vs 0.742 ± 0.091, P < 0.001). Furthermore, according to the results of the MTT assay, SOX11 promoted the proliferation of BMSC (1.064 ± 0.093 vs 0.747 ± 0.090, P < 0.001).ConclusionMiR‐141 inhibited the proliferation of BMSC in SANFH by targeting SOX11. Inhibition of miR‐141 upregulated the expression of SOX11 and promoted the proliferation of BMSC. MiR‐141 and SOX11 could be new targets for investigating the mechanism of SANFH.

Highlights

Glucocorticoids are used in the treatment of many inflammatory, allergic, immunologic, and malignant disorders
MiR-141 inhibited the proliferation of bone marrow mesenchymal stem cells (BMSC) in Steroid-induced avascular necrosis of the femoral head (SANFH) by targeting SOX11
Inhibition of miR-141 upregulated the expression of SOX11 and promoted the proliferation of BMSC

Summary

Introduction

Glucocorticoids are used in the treatment of many inflammatory, allergic, immunologic, and malignant disorders. Steroid-induced avascular necrosis of the femoral head (SANFH) can be induced by the long-term use of glucocorticoids[1,2]. It is well known that long-term use of glucocorticoids leads to abnormal lipid distribution. One possible mechanism for SANFH involves alterations in circulating lipids with resultant microemboli in the arteries supplying bone[4]. Another is the increasing bone marrow adipocyte size and number, which may be able to block the venous outflow[5]. Several studies argue that increased sclerostin production through osteocytes could inhibit long-term bone formation, resulting in suppression of the Wnt/β-catenin pathway and an increase of the peroxisome proliferator-activated receptor.

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