Peng Jun, Zhan Yulin, Zong Yang
Department of Orthopedics, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Cell Tissue Res. 2022 May;388(2):301-312. doi: 10.1007/s00441-022-03588-y. Epub 2022 Feb 22.
N6-methyladenosine (m6A) modification plays a crucial role in the progression of osteoporosis (OP). The study aimed to explore the effects of methyltransferase-like 3 (METTL3) in OP. The levels of METTL3, LINC00657, miR-144-3p and BMPR1B were detected using qPCR. Osteogenesis was assessed using alizarin red and alkaline phosphatase (ALP) staining assays. The protein expression of Bglap, Runx2 and Col1a1 was measured by western blot. The targets of LINC00657 and miR-144-3p were screened by bioinformatic analysis. The interaction between miR-144-3p and LINC00657 or BMPR1B was analyzed by dual-luciferase reporter assay and RNA pull-down assay. The results showed that METTL3 was downregulated in OP. METTL3 mediated m6A methylation of LINC00657 to promote the development of osteogenesis. Further study indicated that LINC00657 functioned as a ceRNA to upregulate BMPR1B via sponging miR-144-3p. Additionally, BMPR1B knockdown alleviated the effects of METTL3 on osteogenesis of bone marrow mesenchymal stem cells (BMSCs). Taken together, METTL3 facilitated osteogenic differentiation of BMSCs via the LINC00657/miR-144-3p/BMPR1B axis. Our findings may provide a novel insight of m6A methylation in the development of OP.
N6-甲基腺苷（m6A）修饰在骨质疏松症（OP）进展中起关键作用。本研究旨在探讨甲基转移酶样3（METTL3）在OP中的作用。采用qPCR检测METTL3、LINC00657、miR-144-3p和BMPR1B的水平。使用茜素红和碱性磷酸酶（ALP）染色试验评估成骨作用。通过蛋白质印迹法检测Bglap、Runx2和Col1a1的蛋白表达。通过生物信息学分析筛选LINC00657和miR-144-3p的靶标。通过双荧光素酶报告基因试验和RNA下拉试验分析miR-144-3p与LINC00657或BMPR1B之间的相互作用。结果显示，OP中METTL3表达下调。METTL3介导LINC00657的m6A甲基化以促进成骨作用的发展。进一步研究表明，LINC00657作为竞争性内源RNA（ceRNA），通过海绵吸附miR-144-3p上调BMPR1B。此外，敲低BMPR1B可减轻METTL3对骨髓间充质干细胞（BMSCs）成骨作用的影响。综上所述，METTL3通过LINC00657/miR-144-3p/BMPR1B轴促进BMSCs的成骨分化。我们的研究结果可能为OP发生发展过程中m6A甲基化提供新的见解。
