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ABSTRACT
Osteoporosis poses a threat to human health worldwide. To date, there have been few studies regarding targeted treatment of osteoporosis. We aimed to identify the possible molecular mechanism of circular RNA (circ)_0062582 in osteogenic differentiation, and the interactions among circ_0062582, microRNA-145 (miR-145) and core-binding factor subunit β (CBFB). The proliferation of human bone marrow mesenchymal stem cells (hBMSCs) was tested with a cell counting kit-8 assay. Circ_0062582, miR-145 and CBFB were overexpressed by transient transfection. Dual-luciferase reporter assay system was used to analyze the combination among circ_0062582, miR-145 and CBFB. Additionally, the levels of circ_0062582, miR-145, CBFB, osterix (OSX), osteocalcin (OCN) and collagen type 1 (COL1) were detected by means of RT-qPCR or western blot analysis. Alkaline phosphatase and Alizarin red stainings were performed to analyze the degree of osteogenic differentiation under the control of circ_0062582, miR-145 and CBFB. The results demonstrated that circ_0062582 level was notably elvated during osteogenic differentiation of hBMSCs. Circ_0062582 overexpression significantly promoted osteogenic differentiation and upregulated the levels of osteogenic differentiation-related proteins, including OSX, OCN and COL1. In addition, miR-145, which was identified as the target gene of circ_0062582, could specifically target CBFB 3′-UTR regions. Next, these changes caused by the overexpression of circ_0062582 were reversed following the addition of miR-145 mimic. Following overexpression of CBFB, osteogenic differentiation was increased. In summary, these results demonstrated that the role of circ_0062582 in osteoporosis is mediated through regulating the expression level of CBFB via miR-145.
GRAPHICAL ABSTRACT
Highlights
1. Circ_0062582 expression was upregulated during osteogenic differentiation of hBMSCs.
2. Circ_0062582 acts as a sponge of miR-145 in hBMSCs.
3. CBFB is a target of miR-145 in hBMSCs, and positively regulated by circ_0062582.
4. Circ_0062582 promotes differentiation of hBMSCs by regulating miR-145/CBFB axis.
Availability of data and materials
All data generated or analyzed during this study are included in this published article.
Disclosure statement
No potential conflict of interest was reported by the author(s).
Ethics approval and consent to participate
The study was approved by the Ethics Committee of The First People’s Hospital of Yancheng (Yancheng, China).
Supplementary material
Supplemental data for this article can be accessed here.
