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

Abstract

Objective To 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). Methods Bone 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. Result The 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). Conclusion MiR‐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.

MicroRNA‑141 inhibits the differentiation of bone marrow‑derived mesenchymal stem cells in steroid‑induced osteonecrosis via E2F3

“…A previous study has indicated that miRNA plays an crucial role in the osteogenic differentiation of BMSCs, and abnormal expression of miRNA molecules may affect this process ( 46 ). Our previous study revealed that miR-141 can inhibit proliferation in BMSCs ( 24 ). Nevertheless, the role of this miRNA in osteogenic differentiation is not clear.…”

“…miR-141 is also associated with tissue repair and osteogenic differentiation ( 23 ). Indeed, miR-141 inhibits the proliferation of BMSCs; inhibition of miR-141 can promote the proliferation of BMSCs ( 24 ) and their osteogenic differentiation ( 5 ), possibly by targeting vitamin C transporter 2 ( 25 ). Therefore, the inhibition of miR-141 could be harnessed for the prevention and treatment of osteonecrosis caused by glucocorticoids.…”

Static Magnetic Fields Enhance the Chondrogenesis of Mandibular Bone Marrow Mesenchymal Stem Cells in Coculture Systems

“…Derived from 3-week-old SD rats (male,70 g, purchased from the Model Animal Research Center of Kunming Medical University), the mandibles ( Figure 1(c) ) were removed and the medullary cavity was exposed [ 28 ]. The medullary cavity was rinsed out using a 1 ml syringe loaded with the SD rat BMSC culture medium (Cyagen Biosciences, USA), and the rinse solution was transferred to a culture flask [ 29 , 30 ].…”

The miR151 and miR5100 Transfected Bone Marrow Stromal Cells Increase Myoblast Fusion in IGFBP2 Dependent Manner

“…They are known to regulate many different biological processes and play an important roles in cell proliferation, migration, and differentiation [ 25 ]. For example, miR141 was shown to inhibit rat BMSC proliferation in vitro by direct targeting of SRY-Box Transcription factor 11 (SOX11) [ 26 ]. miR19b-3p overexpression induces human BMSC proliferation and expression of osteogenic differentiation related proteins: Runt-related transcription factor 2 (RUNX2), collagen type 1, alpha 1 (COL1), and alkaline phosphatase (ALP) [ 26 ].…”

“…For example, miR141 was shown to inhibit rat BMSC proliferation in vitro by direct targeting of SRY-Box Transcription factor 11 (SOX11) [ 26 ]. miR19b-3p overexpression induces human BMSC proliferation and expression of osteogenic differentiation related proteins: Runt-related transcription factor 2 (RUNX2), collagen type 1, alpha 1 (COL1), and alkaline phosphatase (ALP) [ 26 ]. miR335-5p promotes osteogenic differentiation by regulating Dickkopf-related protein 1 (DKK1), a known inhibitor of WNT signaling and osteogenic differentiation [ 27 ].…”