LncRNA FGD5-AS1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells by regulating miR-93-5p/BMP2 axis

doi:10.16352/j.issn.1001-6325.2023.08.1179

Abstract

Abstract: Objective To investigate the impact of long non-coding RNA (lncRNA) FGD5-AS1 on the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) through regulation of the microRNA miR-93-5p/bone morphogenetic protein-2(BMP2) axis. Methods hBM-MSCs in logarithmic growth phase were taken, and the expression levels of lncRNA FGD5-AS1, miR-93-5p and BMP2 mRNA were detected before and after osteogenic differentiation; The cells were transfected or co-transfected with pcDNA FGD5-AS1, miR-93-5p inhibitor, miR-93-5p mimics and corresponding negative controls, respectively, then divided into pcDNA NC group, pcDNA FGD5-AS1 group, inhibitor NC group, miR-93-5p inhibitor group, pcDNA FGD5-AS1+mimics NC group, or pcDNA FGD5-AS1+miR-93-5p mimics group and non-transfected cells were taken as blank group. CCK-8 assay was applied to detect cell proliferation ability of each group; Alkaline phosphatase (ALP) kit was applied to detect its activity; Alizarin red staining was applied to identify cellular mineralized nodule formation; Western blot was applied to detect the levels of BMP2, osteogenesis-related markers-osteocalcin (OCN), osteopontin (OPN), and osterix(OSX); Dual-luciferase experiment was applied to verify the targeting relationship of miR-93-5p with FGD5-AS1 and BMP2, respectively. Results lncRNA FGD5-AS1 and BMP2 were found to be targets of miR-93-5p. After osteogenic differentiation, the expression of FGD5-AS1 and BMP2 was increased and the expression of miR-93-5p was decreased (P＜0.05). Compared with pcDNA NC group, the expression of FGD5-AS1 in pcDNA FGD5-AS1 group was significantly increased(P＜0.05), indicating successful transfection; Mineralized nodules, cell proliferation, ALP activity and the expression of BMP2, OCN, OPN and OSX were obviously higher (P＜0.05) in pcDNA FGD5-AS1 group. Compared with the inhibitor NC group, the expression of miR-93-5p in miR-93-5p inhibitor group was significantly decreased (P＜0.05), indicating successful transfection; Mineralized nodules, cell proliferation, ALP activity and the expression of BMP2, OCN, OPN and OSX were obviously improved (P＜0.05) in miR-93-5p inhibitor group. Over-expression of miR-93-5p inhibited the promoting effect of FGD5-AS1 on the osteogenic differentiation of hBM-MSCs(P＜0.05). Conclusions Up-regulation of FGD-AS1 promotes the osteogenic differentiation of hBM-MSCs, which might be related to the miR-93-5p/BMP2 axis.

Key words: human bone marrow-derived mesenchymal stem cells, lncRNA FGD5-AS1, miR-93-5p/BMP2 axis, osteogenic differentiation

CLC Number:

R394.2

ZHANG Liangliang, ZHAO Chengjin, ZHOU Yuhu, CAO Bo, DUAN Mingming, FENG Yangyang. LncRNA FGD5-AS1 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells by regulating miR-93-5p/BMP2 axis[J]. Basic & Clinical Medicine, 2023, 43(8): 1179-1185.

References 14

[1]	谢兴文, 钟建春, 李鼎鹏,等. 薯蓣皂苷防治原发性骨质疏松的研究进展[J]. 基础医学与临床, 2022, 42:960-963.
[2]	Tian F, Yang HT, Huang T, et al. Involvement of CB2 signalling pathway in the development of osteoporosis by regulating the proliferation and differentiation of hBMSCs[J]. J Cell Mol Med, 2021, 25:2426-2435.
[3]	Asila A, Yang X, Kaisaer Y, et al. SNHG16/miR-485-5p/BMP7 axis modulates osteogenic differentiation of human bone marrow-derived mesenchymal stem cells[J]. J Gene Med,2021, 23:e3296. doi: 10.1002/jgm.3296.
[4]	Li C, Lin X, Zhang C, et al. Long non-coding RNA FGD5-AS1 enhances osteosarcoma cell proliferation and migration by targeting miR-506-3p/RAB3D axis[J]. Hum Cell. 2021, 34:1255-1265.
[5]	Kelch S, Balmayor ER, Seeliger C, et al. MiRNAs in bone tissue correlate to bone mineral density and circulat-ing miRNAs are gender independent in osteoporotic patients[J]. Sci Rep, 2017, 7:15861. doi: 10.1038/s41598-017-16113-x.
[6]	Wang CG, Liao Z, Xiao H, et al. LncRNA KCNQ1OT1 promoted BMP2 expression to regulate osteogenic differentiation by sponging miRNA-214[J]. Exp Mol Pathol, 2019, 107:77-84.
[7]	李丹, 孙文艳, 李晶,等. 老年骨质疏松患者骨折与骨代谢标志物的相关性研究[J]. 临床医药实践, 2015, 24:500-504.
[8]	Yu H, Li Y, Tang J, et al. Long non-coding RNA RP11-84C13.1 promotes osteogenic differentiation of bone mesenchymal stem cells and alleviates osteoporosis progression via the miR-23b-3p/RUNX2 axis[J]. Exp Ther Med, 2021, 22:1340. doi: 10.3892/etm.2021.10775.
[9]	Li L, Zhou X, Zhang JT, et al. Exosomal miR-186 derived from BMSCs promote osteogenesis through hippo signaling pathway in postmenopausal osteoporosis[J]. J Orthop Surg Res, 2021, 16:23. doi: 10.1186/s13018-020-02160-0.
[10]	叶志华, 付金伦, 桂定文,等. 下调长链非编码RNA FGD5-AS1抑制肾癌细胞增殖和侵袭的机制研究[J]. 国际医药卫生导报, 2021, 27:2969-2972.
[11]	Yang Y, Sun Z, Liu F, et al. FGD5-AS1 inhibits osteoarthritis development by modulating miR-302d-3p/TGFBR2 axis[J]. Cartilage, 2021, 13:1412S-1420S.
[12]	郭秀珍, 高斌礼, 郭文,等. LncRNA FGD5-AS1靶向miR-103a-3p对IL-1β诱导的关节软骨细胞凋亡的机制研究[J]. 中国骨质疏松杂志, 2020, 26:832-837.
[13]	Qiao L, Li CG, Liu D. CircRNA_0048211 protects postmenopausal osteoporosis through targeting miRNA-93-5p to regulate BMP2[J]. Eur Rev Med Pharmacol Sci, 2020, 24:3459-3466.
[14]	Zhou R, Miao S, Xu J, et al. Circular RNA circ_0000020 promotes osteogenic differentiation to reduce osteoporosis via sponging microRNA miR-142-5p to up-regulate bone morphogenetic protein BMP2[J]. Bioengineered, 2021, 12:3824-3836.