Rbm15 mediated m6A RNA modification participates in the regulation of AKAP2 on chondrogenesis

Abstract

AbstractPurpose:To investigate the function of AKAP2 in the pathogenesis of AIS by regulating the growth of chondrocyteand explore the mechanism involved.Methods:ATDC-5 cells was used as chondrogenic cell model, and the AKAP2 overexpression and knockdown plasmids were transinfected separately. The proliferation of the cells was tested with EdU and CCK-8 experiments, and the apoptosis was examined by flow cytometry. Subsequently, OE-NC and OE-AKAP2 were selected for RNA-seq and MeRIP-seq analyses to find differently expressed genes and RNA methylation peaks. . The expression of m6A methylases in OE-NC and OE-AKAP2 was then analyzed using a t-test. GSEA and KEGG enrichment analyses were performed to find out the concerning biological process and signals. The results of RNA-seq and MeRIP-seq were combined to seek the functional pathway.Results:AKAP2 knockdown significantly decreased the proliferation of ATDC-5 cells and increased the apoptosis rate, while the overexpression of AKAP2 functioned oppositely. A total of 1216 differentially expressed genes and 1193 differentially expressed peak genes were obtained in OE-AKAP2 group compared with OE-NC, and the expression of Rbm15b was significantly upregulated. Joint analysis of RNA-seq and MeRIP-seq showed that Rbm15b was significantly positively correlated with the cartilage-related gene CCN1. Moreover, GSEA and KEGG enrichment analyses showed that the WNT and NF-κB signaling pathways were significantly correlated to the overexpression of AKAP2 in ATDC-5 cells.Conclusion:Our findings suggest that AKAP2 overexpression promotes cartilage growth and development probably through upregulating Rbm15b in ATDC-5 cells and affecting the expression of CCN1, an important transcription factor in the WNT signaling pathway. This may account for the pathogenicity of AKAP2 variants in AIS to a certain degree.