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Aryl hydrocarbon receptor is critical for both AR-dependent and AR-indifferent enzalutamide resistance in castration-resistant prostate cancer

Oncogene volume 45, pages 1312–1323 (2026)Cite this article

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Abstract

The introduction of next-generation androgen receptor signaling inhibitors (ARSIs) like enzalutamide (ENZ), has improved the clinical management of castration-resistant prostate cancer (CRPC). However, acquired resistance to these therapies often develops rapidly, and the underlying resistance mechanisms remain largely unclear. Here, we identified the aryl hydrocarbon receptor (AHR) as a crucial operator of ENZ-resistant CRPC. AHR is upregulated in three ENZ-resistant human CRPC cell lines (C4-2BENZR, CWR-R1ENZR, and VCaPENZR) as well as in high-grade prostate tumors from patients receiving ENZ treatment. Stable knockdown of AHR substantially reduced the growth of ENZ-resistant CRPC cells and xenografts. Mechanistically, AHR engages in distinct transcriptional programs in a cellular context-dependent manner. AHR directly regulates the transcription and expression of androgen receptor (AR)/glucocorticoid receptor (GR) co-target genes in CWR-R1ENZR cells, suggesting an AR-dependent mechanism of ENZ resistance. AHR promotes neuroendocrine differentiation while suppressing the expression of AR/GR targets in C4-2BENZR cells, indicating an AR-indifferent mechanism of ENZ resistance. The diverse mechanisms triggered by ENZ were also manifested in clinical samples. Collectively, these findings characterize AHR’s contribution to ENZ resistance in CRPC and illuminate the potential of targeting AHR for treating ARSI-resistant advanced prostate cancer.

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Fig. 1: AHR expression is elevated in ENZ-resistant and treated CRPC.
Fig. 2: AHR maintains the growth of ENZ-resistant CRPC cells.
Fig. 3: AHR silencing impedes ENZ-resistant CRPC xenograft growth.
Fig. 4: The canonical AHR-dependent xenobiotic pathway is not responsible for the growth of ENZR CRPC.
Fig. 5: AHR regulates AR/GR target genes in CWR-R1ENZR cells.
Fig. 6: AHR mediates NE differentiation in C4-2BENZR cells.
Fig. 7: ENZ elicits diverse responses in clinical samples.

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Data availability

All sequencing results are available in the NCBI GEO database under the following accession numbers: GSE289313 for AHR ChIP-seq of control and AHR-knockdown CWR-R1ENZR cells, GSE289910 for RNA-seq of control and AHR-knockdown CWR-R1ENZR and C4-2BENZR cells, and GSE290016 for RNA-seq of ENZR and ENZS C4-2B, CWR-R1, and VCaP cells.

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Acknowledgements

We thank Jing Wei for technical help and Gary Mawyer for editorial assistance. This work was supported by NIH/NCI grants R37CA233658, R01CA258634, and R01CA279528 (to BJW).

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy and Pharmaceutical Sciences, Washington State University, Spokane, WA, USA

    Chia-Hui Chen & Boyang Jason Wu

  2. Department of Pathology, The University of Illinois, Chicago, IL, USA

    Ryan Brown & Donald J. Vander Griend

  3. Department of Urologic Surgery, University of California, Davis, Sacramento, CA, USA

    Allen C. Gao

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  1. Chia-Hui Chen
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  2. Ryan Brown
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  3. Donald J. Vander Griend
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  4. Allen C. Gao
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  5. Boyang Jason Wu
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Contributions

BJW and CHC conceptualized the study. CHC performed all experiments, analyzed data, and wrote the draft of the manuscript. BJW designed experiments, edited the manuscript, and acquired funding. RB, DVG, and ACG provided ENZS and ENZR cell lines.

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Correspondence to Boyang Jason Wu.

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All methods were performed in accordance with the relevant guidelines and regulations. All animal studies received prior approval from the Institutional Animal Care and Use Committee (IACUC) at Washington State University (No. 6635) and complied with IACUC recommendations.

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Chen, CH., Brown, R., Vander Griend, D.J. et al. Aryl hydrocarbon receptor is critical for both AR-dependent and AR-indifferent enzalutamide resistance in castration-resistant prostate cancer. Oncogene 45, 1312–1323 (2026). https://doi.org/10.1038/s41388-026-03723-x

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