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

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.