{"title":"METTL3-mediated m6A modification of circSTAT6 modulates miR-188-3p/Beclin1 axis to promote osteogenic differentiation of mesenchymal stem cells.","authors":"Yue Luo, Yubo Shi, Yanqing Wu, Hui Cao","doi":"10.1186/s13018-025-05720-4","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The role of N6-methyladenosine (m6A)-modified circRNAs in disease progression is of great significance. However, the specific impact of m6A modification of circSTAT6 on osteoporosis (OP) is still uncertain.</p><p><strong>Methods: </strong>The qRT-PCR was employed to assess the levels of METTL3, circSTAT6, miR-188-3p, and Beclin1. To investigate the interaction between miR-188-3p and circSTAT6 or Beclin, a dual-luciferase reporter assay was performed. To evaluate osteogenic differentiation in bone marrow mesenchymal stem cell (BMSC), western blot analysis was conducted to evaluate the protein expression of osteogenic markers, including ALP, OPN, and Runx2. In addition, alizarin red and alkaline phosphatase (ALP) staining assays were employed to assess osteogenesis.</p><p><strong>Results: </strong>The findings revealed that the downregulation of circSTAT6 was observed in OP. On the other hand, the overexpression of circSTAT6 was found to enhance the osteogenic differentiation of BMSC. In addition, the involvement of METTL3 in mediating m6A methylation of circSTAT6 was identified, which ultimately promoted osteogenesis. Furthermore, circSTAT6 functioned as an miR-188-3p sponge to regulate the expression of Beclin1. Further study revealed that the osteogenic-enhancing effect caused by circSTAT6 overexpression was counteracted by introducing a miR-188-3p mimic. Similarly, the osteogenic-promoting impact of the miR-188-3p inhibitor was reversed by suppressing Beclin1 expression.</p><p><strong>Conclusions: </strong>The present study revealed, for the first time, that METTL3-mediated m6A modification of circSTAT6 regulated the miR-188-3p/Beclin1 axis to promote the osteogenic differentiation of BMSC. These findings offer a potential therapeutic target for the treatment of OP.</p>","PeriodicalId":16629,"journal":{"name":"Journal of Orthopaedic Surgery and Research","volume":"20 1","pages":"313"},"PeriodicalIF":2.8000,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11938739/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Orthopaedic Surgery and Research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s13018-025-05720-4","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ORTHOPEDICS","Score":null,"Total":0}
引用次数: 0
Abstract
Background: The role of N6-methyladenosine (m6A)-modified circRNAs in disease progression is of great significance. However, the specific impact of m6A modification of circSTAT6 on osteoporosis (OP) is still uncertain.
Methods: The qRT-PCR was employed to assess the levels of METTL3, circSTAT6, miR-188-3p, and Beclin1. To investigate the interaction between miR-188-3p and circSTAT6 or Beclin, a dual-luciferase reporter assay was performed. To evaluate osteogenic differentiation in bone marrow mesenchymal stem cell (BMSC), western blot analysis was conducted to evaluate the protein expression of osteogenic markers, including ALP, OPN, and Runx2. In addition, alizarin red and alkaline phosphatase (ALP) staining assays were employed to assess osteogenesis.
Results: The findings revealed that the downregulation of circSTAT6 was observed in OP. On the other hand, the overexpression of circSTAT6 was found to enhance the osteogenic differentiation of BMSC. In addition, the involvement of METTL3 in mediating m6A methylation of circSTAT6 was identified, which ultimately promoted osteogenesis. Furthermore, circSTAT6 functioned as an miR-188-3p sponge to regulate the expression of Beclin1. Further study revealed that the osteogenic-enhancing effect caused by circSTAT6 overexpression was counteracted by introducing a miR-188-3p mimic. Similarly, the osteogenic-promoting impact of the miR-188-3p inhibitor was reversed by suppressing Beclin1 expression.
Conclusions: The present study revealed, for the first time, that METTL3-mediated m6A modification of circSTAT6 regulated the miR-188-3p/Beclin1 axis to promote the osteogenic differentiation of BMSC. These findings offer a potential therapeutic target for the treatment of OP.
期刊介绍:
Journal of Orthopaedic Surgery and Research is an open access journal that encompasses all aspects of clinical and basic research studies related to musculoskeletal issues.
Orthopaedic research is conducted at clinical and basic science levels. With the advancement of new technologies and the increasing expectation and demand from doctors and patients, we are witnessing an enormous growth in clinical orthopaedic research, particularly in the fields of traumatology, spinal surgery, joint replacement, sports medicine, musculoskeletal tumour management, hand microsurgery, foot and ankle surgery, paediatric orthopaedic, and orthopaedic rehabilitation. The involvement of basic science ranges from molecular, cellular, structural and functional perspectives to tissue engineering, gait analysis, automation and robotic surgery. Implant and biomaterial designs are new disciplines that complement clinical applications.
JOSR encourages the publication of multidisciplinary research with collaboration amongst clinicians and scientists from different disciplines, which will be the trend in the coming decades.