METTL3-mediated m6A modification of circSTAT6 modulates miR-188-3p/Beclin1 axis to promote osteogenic differentiation of mesenchymal stem cells.

IF 2.8 3区 医学 Q1 ORTHOPEDICS
Yue Luo, Yubo Shi, Yanqing Wu, Hui Cao
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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.

mettl3介导的m6A修饰circSTAT6调节miR-188-3p/Beclin1轴促进间充质干细胞成骨分化。
背景:n6 -甲基腺苷(m6A)修饰的环状rna在疾病进展中的作用具有重要意义。然而,m6A修饰circSTAT6对骨质疏松症(OP)的具体影响尚不确定。方法:采用qRT-PCR检测METTL3、circSTAT6、miR-188-3p和Beclin1的表达水平。为了研究miR-188-3p与circSTAT6或Beclin之间的相互作用,我们进行了双荧光素酶报告基因实验。为了评估骨髓间充质干细胞(bone marrow mesenchymal stem cell, BMSC)的成骨分化,我们采用western blot分析方法评估成骨标志物ALP、OPN、Runx2的蛋白表达。此外,采用茜素红和碱性磷酸酶(ALP)染色法评估成骨情况。结果:在op中观察到circSTAT6的下调,另一方面发现circSTAT6的过表达增强了BMSC的成骨分化。此外,METTL3参与介导circSTAT6的m6A甲基化,最终促进成骨。此外,circSTAT6作为miR-188-3p海绵调节Beclin1的表达。进一步的研究表明,引入miR-188-3p模拟物可以抵消circSTAT6过表达引起的成骨增强作用。同样,通过抑制Beclin1的表达,miR-188-3p抑制剂促进成骨的作用被逆转。结论:本研究首次揭示了mettl3介导的m6A修饰circSTAT6调控miR-188-3p/Beclin1轴促进BMSC成骨分化。这些发现为OP的治疗提供了一个潜在的治疗靶点。
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来源期刊
CiteScore
4.10
自引率
7.70%
发文量
494
审稿时长
>12 weeks
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.
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