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Overexpression of androgen receptor enhances the binding of the receptor to the chromatin in prostate cancer

Oncogene volume 31, pages 2153–2163 (2012)Cite this article

Abstract

Androgen receptor (AR) is overexpressed in the majority of castration-resistant prostate cancers (CRPCs). Our goal was to study the effect of AR overexpression on the chromatin binding of the receptor and to identify AR target genes that may be important in the emergence of CRPC. We have established two sublines of LNCaP prostate cancer (PC) cell line, one overexpressing AR 2–3-fold and the other 4–5-fold compared with the control cells. We used chromatin immunoprecipitation (ChIP) and deep-sequencing (seq) to identify AR-binding sites (ARBSs). We found that the number of ARBSs and the AR-binding strength were positively associated with the level of AR when cells were stimulated with low concentrations of androgens. In cells overexpressing AR, the chromatin binding of the receptor took place in 100-fold lower concentration of the ligand than in control cells. We confirmed the association of AR level and chromatin binding in two PC xenografts, one containing AR gene amplification with high AR expression, and the other with low expression. By combining the ChIP-seq and expression profiling, we identified AR target genes that are upregulated in PC. Of them, the expression of ZWINT, SKP2 (S-phase kinase-associated protein 2 (p45)) and FEN1 (flap structure-specific endonuclease 1) was demonstrated to be increased in CRPC, while the expression of SNAI2 was decreased in both PC and CRPC. FEN1 protein expression was also associated with poor prognosis in prostatectomy-treated patients. Finally, the knock-down of FEN1 with small interfering RNA inhibited the growth of LNCaP cells. Our data demonstrate that the overexpression of AR sensitizes the receptor binding to chromatin, thus, explaining how AR signaling pathway is reactivated in CRPC cells.

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Acknowledgements

We wish to thank Ms Mariitta Vakkuri, Ms Päivi Martikainen and Mr Rolle Rahikainen for the skillful technical assistance. The research leading to these results has received funding from the European Union FP6, CANCURE Programme (contract number: MEST-CT-2005-020970). In addition, grant support has been received from Academy of Finland, Cancer Society of Finland, Reino Lahtikari Foundation, Sigrid Juselius Foundation, and the Medical Research Fund of Tampere University Hospital.

Author contributions: TV and AU designed the study, analyzed the data and wrote the paper. AU and BS performed experiments and edited the paper. JS, AL, HL performed the bioinformatic analyses and edited the paper. LML performed experiments and edited the paper. KKW provided the expression data and edited the paper. OAJ contributed new reagents and edited the paper. TLT provided clinical material and edited the paper. RLV provided the xenografts material and edited the paper.

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Authors and Affiliations

  1. Institute of Biomedical Technology, University of Tampere and Tampere University Hospital, Tampere, Finland

    A Urbanucci, L M Latonen, K K Waltering & T Visakorpi

  2. University of Helsinki, Institute of Biomedicine Physiology, Biomedicum Helsinki, Helsinki, Finland

    B Sahu & O A Jänne

  3. Department of Signal Processing, Tampere University of Technology, Tampere, Finland

    J Seppälä & A Larjo

  4. Department of Urology, University of Tampere and Tampere University Hospital, Tampere, Finland

    T L J Tammela

  5. Department of Urology, University of Washington Medical Center, NE Pacific, Seattle, WA, USA

    R L Vessella

  6. Department of Information and Computer Science, Aalto University School of Science and Technology, Helsinki, Finland

    H Lähdesmäki

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  1. A Urbanucci
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Correspondence to T Visakorpi.

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Urbanucci, A., Sahu, B., Seppälä, J. et al. Overexpression of androgen receptor enhances the binding of the receptor to the chromatin in prostate cancer. Oncogene 31, 2153–2163 (2012). https://doi.org/10.1038/onc.2011.401

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