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METTL14 通过调控 pri-miR-873 的加工来介导 m6A 修饰对骨髓间充质干细胞成骨增殖和分化的影响。

METTL14 mediates ma modification on osteogenic proliferation and differentiation of bone marrow mesenchymal stem cells by regulating the processing of pri-miR-873.

机构信息

Department of Orthopedic Surgery, Tangdu Hospital, Air Force Military Medical University, Xi'an, Shaanxi 710038, P.R. China.

出版信息

Mol Med Rep. 2023 Sep;28(3). doi: 10.3892/mmr.2023.13053. Epub 2023 Jul 14.

Abstract

N-methyl-adenosine (m6a) is involved in the occurrence and development of various diseases such as autogenic immune disease and tumors. Methyltransferases regulate primary (pri)-microRNA (miRNA/miR) processing by mediating m6a modifications, consequently affecting pathological processes including immune-related diseases by regulating both innate and adaptive immune cells. However, the roles of m6a on the biological functions of bone marrow mesenchymal stem cells (BMSCs) remain to be elucidated. The relative expression levels of methyltransferase-like 14 (METTL14) and other methyltransferases, demethylases, and miR-873 in bone samples from patients with osteoporosis and from normal individuals were measured by reverse transcription-quantitative PCR. Cell Counting Kit-8 assay was used to examine the proliferation of BMSCs. Co-immunoprecipitation (Co-IP) was used to investigate the binding of METTL14 to DiGeorge syndrome critical region 8 (DGCR8). RNA immunoprecipitation (RIP) was used to examine the binding of METTL14 to pri-miR-873. METTL14 and ma modifications were highly detected in patients with osteoporosis compared with the controls. Co-IP results indicated that silencing of METTL14 reduced METTL14 and ma modification levels in BMSCs. Downregulation of METTL14 significantly promoted the proliferation of BMSCs. RIP results suggested that METTL14/ma methylation modification promoted the processing of pri-miR-873 by binding to DGCR8 in BMSCs. Furthermore, overexpression of miR-873 inhibited the proliferation of BMSCs. The results also showed that miR-873 mimics significantly inhibited the proliferation in small interfering (si)-METTL14 transfected BMSCs; however, miR-873 inhibitors markedly promoted the proliferation of si-METTL14 transfected BMSCs. METTL14 and ma modifications were upregulated in osteoporosis samples. METTL14 promoted the processing of pri-miR-873 into mature miR-873 by regulating ma modification. Furthermore, overexpression of miR-873 significantly inhibited the proliferation of BMSCs. Therefore, the METTL14/ma/miR-873 axis may be a potential target for the treatment of osteoporosis.

摘要

N6-甲基腺苷(m6A)参与自身免疫性疾病和肿瘤等多种疾病的发生和发展。甲基转移酶通过调节 m6A 修饰来调节初级(pri)-微小 RNA(miRNA/miR)的加工,从而通过调节先天和适应性免疫细胞来影响包括免疫相关疾病在内的病理过程。然而,m6A 对骨髓间充质干细胞(BMSCs)的生物学功能的作用仍有待阐明。通过逆转录定量 PCR 测量骨质疏松症患者和正常人骨样本中甲基转移酶样 14(METTL14)和其他甲基转移酶、去甲基酶和 miR-873 的相对表达水平。细胞计数试剂盒-8 测定用于检测 BMSCs 的增殖。免疫共沉淀(Co-IP)用于研究 METTL14 与 DiGeorge 综合征关键区域 8(DGCR8)的结合。RNA 免疫沉淀(RIP)用于检测 METTL14 与 pri-miR-873 的结合。与对照组相比,骨质疏松症患者中 METTL14 和 ma 修饰高度检测到。Co-IP 结果表明,沉默 METTL14 降低了 BMSCs 中的 METTL14 和 ma 修饰水平。下调 METTL14 可显著促进 BMSCs 的增殖。RIP 结果表明,METTL14/ma 甲基化修饰通过与 BMSCs 中的 DGCR8 结合促进 pri-miR-873 的加工。此外,过表达 miR-873 抑制 BMSCs 的增殖。结果还表明,miR-873 模拟物可显著抑制 si-METTL14 转染的 BMSCs 的增殖;然而,miR-873 抑制剂可显著促进 si-METTL14 转染的 BMSCs 的增殖。METTL14 和 ma 修饰在骨质疏松症样本中上调。METTL14 通过调节 ma 修饰促进 pri-miR-873 向成熟 miR-873 的加工。此外,过表达 miR-873 可显著抑制 BMSCs 的增殖。因此,METTL14/ma/miR-873 轴可能是治疗骨质疏松症的潜在靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9ab/10407617/670365564904/mmr-28-03-13053-g00.jpg

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