C-X-C motif chemokine receptor 4 (CXCR4) is highly expressed in aldosterone-producing adenoma, and gallium-68 pentixafor PET-CT imaging targeting CXCR4 has been utilized for subtype diagnosis in primary aldosteronism. However, the roles of CXCR4 in regulating aldosterone biosynthesis remain poorly understood. In this study, we observed a strong co-localization of aldosterone synthase (CYP11B2) and CXCR4 in aldosterone-producing adenoma and other aldosterone-producing lesions. Functional experiments in H295R cells revealed that CXCR4 overexpression significantly suppressed both aldosterone synthesis and CYP11B2 expression, whereas CXCR4 knockdown conversely enhanced aldosterone production and up-regulated CYP11B2. Mechanistically, CXCR4 inhibited aldosterone biosynthesis by up-regulating inhibitor of DNA binding (ID) proteins, which directly repressed CYP11B2 transcription. Our data demonstrate that in aldosterone-producing lesions, CXCR4 expression is consistently elevated alongside CYP11B2, indicating a potential compensatory mechanism to counteract elevated aldosterone levels.