m⁶A介导的内分泌治疗耐药乳腺癌激酶重编程的调控

Endocrine therapy resistance (ETR) in estrogen receptor-positive breast cancer arises through multiple adaptive mechanisms, with kinase reprogramming representing a central driver of therapeutic escape. N 6 -methyladenosine (m 6 A), the most abundant internal RNA modification, has emerged as a key regulator of RNA fate, yet its role in restructuring kinase networks during ETR is unclear. Using GLORI sequencing, a single-nucleotide-resolution method for stoichiometric m 6 A quantification, we profiled parental, tamoxifen and exemestane resistant MCF7 derivatives. Tamoxifen-resistant cells exhibited ~2,600 m 6 A sites with canonical DRACH motifs with widespread hyper- and hypomethylation across transcripts encoding MAPK regulators, receptor tyrosine kinases, and cell-cycle-associated kinases. Multi-omic integration including RNA-seq and quantitative proteomics indicate differentially methylated kinase mRNAs gained stability, enhanced translational efficiency and increased protein output, establishing an m 6 A-dependent mechanism of kinase reprogramming. Resistant cells also displayed selective upregulation of m 6 A-associated RNA-binding proteins, including YTHDF1 and IGF2BP2, consistent with a remodeled post-transcriptional landscape. Together, our findings uncover a previously underappreciated epitranscriptomic layer integrating quantitative m 6 A dynamics and RNA-binding protein shifts that drives kinase reprogramming and promotes ETR in breast cancer.