Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression

Oncogene volume 31, pages 4759–4767 (2012)Cite this article

Subjects

Abstract

Reactivation of the androgen receptor (AR) during androgen depletion therapy (ADT) underlies castration-resistant prostate cancer (CRPCa). Alternative splicing of the AR gene and synthesis of constitutively active COOH-terminally truncated AR variants lacking the AR ligand-binding domain has emerged as an important mechanism of ADT resistance in CRPCa. In a previous study, we demonstrated that altered AR splicing in CRPCa 22Rv1 cells was linked to a 35-kb intragenic tandem duplication of AR exon 3 and flanking sequences. In this study, we demonstrate that complex patterns of AR gene copy number imbalances occur in PCa cell lines, xenografts and clinical specimens. To investigate whether these copy number imbalances reflect AR gene rearrangements that could be linked to splicing disruptions, we carried out a detailed analysis of AR gene structure in the LuCaP 86.2 and CWR-R1 models of CRPCa. By deletion-spanning PCR, we discovered a 8579-bp deletion of AR exons 5, 6 and 7 in the LuCaP 86.2 xenograft, which provides a rational explanation for synthesis of the truncated AR v567es AR variant in this model. Similarly, targeted resequencing of the AR gene in CWR-R1 cells led to the discovery of a 48-kb deletion in AR intron 1. This intragenic deletion marked a specific CWR-R1 cell population with enhanced expression of the truncated AR-V7/AR3 variant, a high level of androgen-independent AR transcriptional activity and rapid androgen independent growth. Together, these data demonstrate that structural alterations in the AR gene are linked to stable gain-of-function splicing alterations in CRPCa.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  1. Chen Y, Clegg NJ, Scher HI . Anti-androgens and androgen-depleting therapies in prostate cancer: new agents for an established target. Lancet Oncol 2009; 10: 981–991.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Heemers HV, Tindall DJ . Androgen receptor (AR) coregulators: a diversity of functions converging on and regulating the AR transcriptional complex. Endocr Rev 2007; 28: 778–808.

    Article  CAS  PubMed  Google Scholar 

  3. Scher HI, Sawyers CL . Biology of progressive, castration-resistant prostate cancer: directed therapies targeting the androgen-receptor signaling axis. J Clin Oncol 2005; 23: 8253–8261.

    Article  CAS  PubMed  Google Scholar 

  4. Taplin ME . Drug insight: role of the androgen receptor in the development and progression of prostate cancer. Nat Clin Pract Oncol 2007; 4: 236–244.

    Article  CAS  PubMed  Google Scholar 

  5. Attard G, Swennenhuis JF, Olmos D, Reid AH, Vickers E, A’Hern R et al. Characterization of ERG, AR and PTEN gene status in circulating tumor cells from patients with castration-resistant prostate cancer. Cancer Res 2009; 69: 2912–2918.

    Article  CAS  PubMed  Google Scholar 

  6. Chen CD, Welsbie DS, Tran C, Baek SH, Chen R, Vessella R et al. Molecular determinants of resistance to antiandrogen therapy. Nat Med 2004; 10: 33–39.

    Article  PubMed  Google Scholar 

  7. Edwards J, Krishna NS, Grigor KM, Bartlett JM . Androgen receptor gene amplification and protein expression in hormone refractory prostate cancer. Br J Cancer 2003; 89: 552–556.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ford 3rd OH, Gregory CW, Kim D, Smitherman AB, Mohler JL . Androgen receptor gene amplification and protein expression in recurrent prostate cancer. J Urol 2003; 170: 1817–1821.

    Article  CAS  PubMed  Google Scholar 

  9. Linja MJ, Savinainen KJ, Saramaki OR, Tammela TL, Vessella RL, Visakorpi T . Amplification and overexpression of androgen receptor gene in hormone-refractory prostate cancer. Cancer Res 2001; 61: 3550–3555.

    CAS  PubMed  Google Scholar 

  10. Liu W, Laitinen S, Khan S, Vihinen M, Kowalski J, Yu G et al. Copy number analysis indicates monoclonal origin of lethal metastatic prostate cancer. Nat Med 2009; 15: 559–565.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Visakorpi T, Hyytinen E, Koivisto P, Tanner M, Keinanen R, Palmberg C et al. In vivo amplification of the androgen receptor gene and progression of human prostate cancer. Nat Genet 1995; 9: 401–406.

    Article  CAS  PubMed  Google Scholar 

  12. Leversha MA, Han J, Asgari Z, Danila DC, Lin O, Gonzalez-Espinoza R et al. Fluorescence in situ hybridization analysis of circulating tumor cells in metastatic prostate cancer. Clin Cancer Res 2009; 15: 2091–2097.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Haapala K, Hyytinen ER, Roiha M, Laurila M, Rantala I, Helin HJ et al. Androgen receptor alterations in prostate cancer relapsed during a combined androgen blockade by orchiectomy and bicalutamide. Lab Invest 2001; 81: 1647–1651.

    Article  CAS  PubMed  Google Scholar 

  14. Hyytinen ER, Haapala K, Thompson J, Lappalainen I, Roiha M, Rantala I et al. Pattern of somatic androgen receptor gene mutations in patients with hormone-refractory prostate cancer. Lab Invest 2002; 82: 1591–1598.

    Article  CAS  PubMed  Google Scholar 

  15. Steinkamp MP, O’Mahony OA, Brogley M, Rehman H, Lapensee EW, Dhanasekaran S et al. Treatment-dependent androgen receptor mutations in prostate cancer exploit multiple mechanisms to evade therapy. Cancer Res 2009; 69: 4434–4442.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Taplin ME, Bubley GJ, Ko YJ, Small EJ, Upton M, Rajeshkumar B et al. Selection for androgen receptor mutations in prostate cancers treated with androgen antagonist. Cancer Res 1999; 59: 2511–2515.

    CAS  PubMed  Google Scholar 

  17. Taplin ME, Bubley GJ, Shuster TD, Frantz ME, Spooner AE, Ogata GK et al. Mutation of the androgen-receptor gene in metastatic androgen-independent prostate cancer. N Engl J Med 1995; 332: 1393–1398.

    Article  CAS  PubMed  Google Scholar 

  18. Taplin ME, Rajeshkumar B, Halabi S, Werner CP, Woda BA, Picus J et al. Androgen receptor mutations in androgen-independent prostate cancer: Cancer and Leukemia Group B Study 9663. J Clin Oncol 2003; 21: 2673–2678.

    Article  CAS  PubMed  Google Scholar 

  19. Thompson J, Hyytinen ER, Haapala K, Rantala I, Helin HJ, Janne OA et al. Androgen receptor mutations in high-grade prostate cancer before hormonal therapy. Lab Invest 2003; 83: 1709–1713.

    Article  CAS  PubMed  Google Scholar 

  20. Tilley WD, Wilson CM, Marcelli M, McPhaul MJ . Androgen receptor gene expression in human prostate carcinoma cell lines. Cancer Res 1990; 50: 5382–5386.

    CAS  PubMed  Google Scholar 

  21. Mohler JL, Gregory CW, Ford 3rd OH, Kim D, Weaver CM, Petrusz P et al. The androgen axis in recurrent prostate cancer. Clin Cancer Res 2004; 10: 440–448.

    Article  CAS  PubMed  Google Scholar 

  22. Montgomery RB, Mostaghel EA, Vessella R, Hess DL, Kalhorn TF, Higano CS et al. Maintenance of intratumoral androgens in metastatic prostate cancer: a mechanism for castration-resistant tumor growth. Cancer Res 2008; 68: 4447–4454.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Locke JA, Guns ES, Lubik AA, Adomat HH, Hendy SC, Wood CA et al. Androgen levels increase by intratumoral de novo steroidogenesis during progression of castration-resistant prostate cancer. Cancer Res 2008; 68: 6407–6415.

    Article  CAS  PubMed  Google Scholar 

  24. Mohler JL, Titus MA, Bai S, Kennerley BJ, Lih FB, Tomer KB et al. Activation of the androgen receptor by intratumoral bioconversion of androstanediol to dihydrotestosterone in prostate cancer. Cancer Res 2011; 71: 1486–1496.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Attard G, Richards J, de Bono JS . New strategies in metastatic prostate cancer: targeting the androgen receptor signaling pathway. Clin Cancer Res 2011; 17: 1649–1657.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. de Bono JS, Logothetis CJ, Molina A, Fizazi K, North S, Chu L et al. Abiraterone and increased survival in metastatic prostate cancer. N Engl J Med 2011; 364: 1995–2005.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Dehm SM, Schmidt LJ, Heemers HV, Vessella RL, Tindall DJ . Splicing of a novel androgen receptor exon generates a constitutively active androgen receptor that mediates prostate cancer therapy resistance. Cancer Res 2008; 68: 5469–5477.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Guo Z, Yang X, Sun F, Jiang R, Linn DE, Chen H et al. A novel androgen receptor splice variant is up-regulated during prostate cancer progression and promotes androgen depletion-resistant growth. Cancer Res 2009; 69: 2305–2313.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Hu R, Dunn TA, Wei S, Isharwal S, Veltri RW, Humphreys E et al. Ligand-independent androgen receptor variants derived from splicing of cryptic exons signify hormone-refractory prostate cancer. Cancer Res 2009; 69: 16–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Sun S, Sprenger CC, Vessella RL, Haugk K, Soriano K, Mostaghel EA et al. Castration resistance in human prostate cancer is conferred by a frequently occurring androgen receptor splice variant. J Clin Invest 2010; 120: 2715–2730.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Watson PA, Chen YF, Balbas MD, Wongvipat J, Socci ND, Viale A et al. Constitutively active androgen receptor splice variants expressed in castration-resistant prostate cancer require full-length androgen receptor. Proc Natl Acad Sci USA 2010; 107: 16759–16765.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Hornberg E, Ylitalo EB, Crnalic S, Antti H, Stattin P, Widmark A et al. Expression of androgen receptor splice variants in prostate cancer bone metastases is associated with castration-resistance and short survival. PLoS One 2011; 6: e19059.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Li Y, Alsagabi M, Fan D, Bova GS, Tewfik AH, Dehm SM . Intragenic rearrangement and altered RNA splicing of the androgen receptor in a cell-based model of prostate cancer progression. Cancer Res 2011; 71: 2108–2117.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Tarailo-Graovac M, Chen N . Using RepeatMasker to identify repetitive elements in genomic sequences. Curr Protoc Bioinformatics 2009. Chapter 4: Unit 4 10..

  35. Quinlan AR, Clark RA, Sokolova S, Leibowitz ML, Zhang Y, Hurles ME et al. Genome-wide mapping and assembly of structural variant breakpoints in the mouse genome. Genome Res 2010; 20: 623–635.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Gottlieb B, Beitel LK, Wu JH, Trifiro M . The androgen receptor gene mutations database (ARDB): 2004 update. Hum Mutat 2004; 23: 527–533.

    Article  CAS  PubMed  Google Scholar 

  37. Berger MF, Lawrence MS, Demichelis F, Drier Y, Cibulskis K, Sivachenko AY et al. The genomic complexity of primary human prostate cancer. Nature 2011; 470: 214–220.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Robbins CM, Tembe WA, Baker A, Sinari S, Moses TY, Beckstrom-Sternberg S et al. Copy number and targeted mutational analysis reveals novel somatic events in metastatic prostate tumors. Genome Res 2011; 21: 47–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Corey E, Quinn JE, Buhler KR, Nelson PS, Macoska JA, True LD et al. LuCaP 35: a new model of prostate cancer progression to androgen independence. Prostate 2003; 55: 239–246.

    Article  CAS  PubMed  Google Scholar 

  40. Roudier MP, True LD, Higano CS, Vesselle H, Ellis W, Lange P et al. Phenotypic heterogeneity of end-stage prostate carcinoma metastatic to bone. Hum Pathol 2003; 34: 646–653.

    Article  PubMed  Google Scholar 

  41. Dagvadorj A, Tan SH, Liao Z, Cavalli LR, Haddad BR, Nevalainen MT . Androgen-regulated and highly tumorigenic human prostate cancer cell line established from a transplantable primary CWR22 tumor. Clin Cancer Res 2008; 14: 6062–6072.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Gregory CW, Johnson Jr RT, Mohler JL, French FS, Wilson EM . Androgen receptor stabilization in recurrent prostate cancer is associated with hypersensitivity to low androgen. Cancer Res 2001; 61: 2892–2898.

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We are grateful to the Minnesota Supercomputing Institute and the Masonic Cancer Center Biostatistics and Bioinformatics Core for providing computing, bioinformatics, statistical, software and data storage support for this project. Cytogenetic analyses were performed in the Cytogenetics Shared Resource Laboratory at the University of Minnesota, with support from the comprehensive Masonic Cancer Center NIH Grant P30 CA077598. We thank Amanda Hemmingsen Jaeger for assistance with developing MLPA for AR gene structure analysis. SMD is a Masonic Scholar of the Masonic Cancer Center, University of Minnesota. This work was supported by a Young Investigator Award from the Prostate Cancer Foundation (SMD), DOD New Investigator Award PC094384 (SMD) NCI Grant CA141011 (SMD), and a seed grant from the Institute of Human Genetics, University of Minnesota.

Author information

Authors and Affiliations

  1. Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA

    Y Li, T H Hwang, K A Silverstein & S M Dehm

  2. Biostatistics and Bioinformatics Core, Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA

    T H Hwang & K A Silverstein

  3. Cytogenetics Shared Resource, Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA

    L A Oseth & B Hirsch

  4. Biomedical Genomics Center, University of Minnesota, Minneapolis, MN, USA

    A Hauge & K B Beckman

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

    R L Vessella

  6. Puget Sound VA Health Care System, Seattle, WA, USA

    R L Vessella

  7. Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, MN, USA

    S C Schmechel, B Hirsch & S M Dehm

  8. BioNet, University of Minnesota, Minneapolis, MN, USA

    S C Schmechel

  9. Institute of Human Genetics, University of Minnesota, Minneapolis, MN, USA

    B Hirsch

  10. These authors contributed equally to this work.,

    Y Li & T H Hwang

Authors
  1. Y Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T H Hwang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L A Oseth
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Hauge
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R L Vessella
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. S C Schmechel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. B Hirsch
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. K B Beckman
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K A Silverstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. S M Dehm
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to S M Dehm.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, Y., Hwang, T., Oseth, L. et al. AR intragenic deletions linked to androgen receptor splice variant expression and activity in models of prostate cancer progression. Oncogene 31, 4759–4767 (2012). https://doi.org/10.1038/onc.2011.637

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2011.637

Keywords

This article is cited by

Search

Advanced search

Quick links