RBM15-mediated m6A modification upregulates KDM2A to promote ferroptosis in osteoarthritis cells

Abstract

Background and aims

Ferroptosis has been acknowledged to be involved in the progression of osteoarthritis (OA), contributing to chondrocyte damage and articular cartilage degradation. This investigation seeks to explain the mechanism of RBM15 in ferroptosis of OA cell models.

Methods

OA cell model was established using IL-1β-induced chondrocytes. After silencing RBM15 expression, TNF-α and IL-6 were detected, cell viability was measured, LDH, Fe²⁺, ROS, MDA, and GSH were assessed, and ACSL4 and SLC7A11 expression was quantified. RT-qPCR and Western blot were employed to detect the expression of RBM15, KDM2A, and HOXA2. The m6A content in cells was assayed using a kit. RIP assay was utilized to analyze m6A modification on KDM2A. Dual-luciferase assay was employed to validate RBM15-mediated m6A modification on KDM2A. RT-qPCR was applied to examine KDM2A mRNA stability. ChIP was employed to assess the enrichment of KDM2A and H3K36me2 at the HOXA2 promoter. Combined experiments were designed to elucidate the role of KDM2A and HOXA2 in ferroptosis of OA cell models.

Results

RBM15 and KDM2A were upregulated, while HOXA2 was reduced in IL-1β-stimulated chondrocytes. RBM15 downregulation enhanced chondrocyte viability and mitigated ferroptosis. RBM15-mediated m6A modification upregulated KDM2A, suppressed H3K36me2 enrichment at the HOXA2 promoter, and inhibited HOXA2 expression. Overexpression of KDM2A and downregulation of HOXA2 could partially counteract the suppressive effect of RBM15 silencing on chondrocyte ferroptosis.

Conclusion

RBM15-mediated m6A modification upregulates KDM2A to inhibit HOXA2 expression by reducing H3K36me2 enrichment, thus promoting IL-1β-induced chondrocyte ferroptosis.