Acyl-CoA synthetase 4 (ACSL4) is involved in arachidonic acid metabolism and is linked to prostate and breast cancer aggressiveness. In prostate cancer, androgen receptor (AR) inhibition leads to ACSL4 overexpression, suggesting that this enzyme has an important role in hormone-resistant tumor. AR is also being studied in breast cancer (BC), potentially serving as a therapeutic target in AR+ BC. In triple-negative breast cancer (TNBC) AR expression has recently emerged as an important factor. TNBC AR+ is a potential target for AR antagonists, currently under clinical trials. However,70-80% of TNBC lack AR expression (QNBC), resulting in highly aggressive tumors with poor survival and resistance to chemotherapy. This study aims to explore androgenic regulation of ACSL4 in BC cells, evaluating AR and ACSL4 as therapeutic targets. AR-positive cells MCF-7 showed low ACSL4 levels while MDA-MB-231, a QNBC model, exhibited high ACSL4 expression. In this work, an AR consensus site was identified in the ACSL4 promoter. We observed that AR overexpression in MDA-MB-231 reduced ACSL4 promoter activity and mRNA levels. In MCF-7 and MDA-MB-231 cells transiently transfected with AR, dihydrotestosterone treatment decreased ACSL4 promoter activity and mRNA levels, whereas ACSL4 promoter activity was increased by AR antagonist bicalutamide (CDX). In MCF-7 cells AR stable silencing increased ACSL4 mRNA levels and ACSL4 promoter activity was increased by site-directed mutagenesis of the AR element. This regulation seems to be reciprocal, as the ACSL4 inhibitor PRGL493 increased AR mRNA levels in MDA-MB-231 cells. Combining submaximal doses of ACSL4 inhibitors (PRGL493 and Rosiglitazone) with CDX, MDA-MB-231 cell proliferation was synergistically decreased. All these findings indicate that AR reduces ACSL4 expression, and this effect is reciprocal. Our results also suggest the importance of evaluating in BC the effectiveness of therapies that combine ACSL4 and AR as therapeutic targets.