Deregulation of homeobox genes has been associated with several human malignancies. For instance, Distal-less homeobox-1 (DLX1), which is involved in the development of craniofacial features and GABAergic interneuron, is also highly upregulated in prostate cancer. We showed that ~60% of advanced-stage prostate cancer patients display higher DLX1 levels, and are associated with metastatic disease and poor survival. We established the oncogenic role of DLX1 in prostate cancer, and deciphered its transcriptional regulation involving ERG, AR/AR-V7 and FOXA1. Moreover, we showed that BET inhibitor and/or anti-androgen drugs disrupt ERG/AR-mediated DLX1 transcription leading to its reduced expression and downstream oncogenic effects. Taken together, we offered a strategy to include BET inhibitors and anti-androgens for the treatment of DLX1-positive prostate cancer patients.