DNMT1-DUOXA1 axis discovers a novel methylation-ferroptosis circuit in bicalutamide-resistant prostate cancer

Prostate cancer, a prevalent malignancy in the male reproductive system, poses significant therapeutic challenges due to the development of resistance to androgen deprivation therapies such as bicalutamide. While current research predominantly focuses on androgen receptor (AR)-dependent mechanisms of resistance, non-AR-dependent pathways remain poorly understood. Here, we report a novel non-AR-dependent mechanism of bicalutamide resistance centered on DUOXA1, a maturation factor for dual oxidases (DUOX) that catalyzes hydrogen peroxide (H 2 O 2 ) production. Our analysis of RNA sequencing data from bicalutamide-resistant and sensitive prostate cancer cells revealed DUOXA1 as a significantly downregulated gene in resistant cells. We demonstrate that hypoxia in the prostate cancer microenvironment enhances HIF1α transcriptional activity, leading to increased DNMT1 expression. DNMT1, an epigenetic modifier, mediates the methylation of the DUOXA1 promoter, thereby silencing its expression. This epigenetic silencing of DUOXA1 inhibits ferroptosis, a form of regulated cell death characterized by iron metabolism disruption and lipid peroxidation, thereby promoting bicalutamide resistance. Our findings indicate that DUOXA1 can enhance bicalutamide resistance by promoting ferroptosis through ROS generation. This study not only provides mechanistic insights into the role of DUOXA1 in bicalutamide resistance but also highlights the HIF1α-DNMT1-DUOXA1 axis as a critical regulator of resistance. Our work suggests potential therapeutic strategies to overcome resistance through epigenetic modulation and activation of DUOXA1, offering a novel perspective on the molecular mechanisms of bicalutamide resistance and paving the way for the development of improved treatment approaches for advanced prostate cancer.