Identification of m6A-Associated mRNAs, lncRNAs, and m6A Regulators in Endometriosis

BACKGROUND: Endometriosis (EMs), which causes significant pain and infertility, continues to affect women of reproductive age, yet its molecular mechanisms remain inadequately defined. N6-methyladenosine (m6A) modification is the most prevalent post-transcriptional modification in mammalian mRNA, and recent studies have increasingly focused on elucidating the relationship between m6A and EMs, to provide theoretical insights for clinical prevention and treatment. METHODS: Using human m6A-mRNA and lncRNA epigenomic microarray techniques, functional and pathway enrichment analysis, protein-protein interaction network (PPI), and R packages, we conducted exploratory analyses to identify mRNAs and lncRNAs with differential methylation, as well as important m6A regulators associated with EMs. These findings were verified using external databases, along with RNA extraction and quantitative real-time PCR (qPCR), immunohistochemistry, and immunofluorescence techniques. RESULTS: Specifically, our analysis revealed 1631 hyper-upregulated, 219 hypo-upregulated, and 6483 hypo-downregulated differentially methylated and expressed genes (DMEGs). Additionally, we identified 8 candidate hub genes-RHOA, KCNA2, BDNF, CACNA1C, FGFR1, IL18, KCNIP2, and RPL17-and characterized their enriched pathways. Furthermore, we observed 6 significantly hypermethylated and 14 significantly hypomethylated lncRNAs in EMs. Our screening identified two key m6A regulators, WTAP and RBM15, which may represent candidate molecules worthy of further investigation in the context of EMs. CONCLUSION: This exploratory screening analysis delineates aberrant expression signatures of m6A regulators and their associated mRNAs and lncRNAs in endometriosis tissues, yielding preliminary molecular profiles that merely reflect correlations rather than causal or definite functional relationships.