Androgen receptor phosphorylation at serine 81 and serine 213 in castrate-resistant prostate cancer



Despite increases in diagnostics and effective treatments, over 300,000 men die from prostate cancer highlighting the need for specific and differentiating biomarkers. AR phosphorylation associates with castrate-resistance, with pARser213 promoting transcriptional activity. We hypothesise that combined pARser81 and pARser213 reduces survival and would benefit from dual-targeting androgen-dependent and Akt-driven disease.


Immunohistochemistry and immunofluorescence were performed on matched hormone-naive and castrate-resistant prostate cancer samples. TempO-Seq gene profiling was analysed using DESeq2 package. LNCaP-AI cells were stimulated with DHT or EGF. WST-1 assays were performed to determine effects of Enzalutamide and BKM120 on cell viability.


Following the development of castrate-resistance, pARser81 expression reduced and pARser213 expression increased. Castrate-resistance pARser81 expression was not associated with survival but high pARser213 expression was associated with reduced survival from relapse. Combined high pARser81 and pARser213 was associated with reduced survival from relapse. pARser81 expression was induced by 10 nM DHT or 10 nM EGF and pARser213 expression was induced by treatment with 10 nM EGF in LNCaP-AI cells. Cell viability was reduced following treatment with 10 nM Enzalutamide and 10 nM BKM120. Eight genes were differentially expressed between hormone-naive and castrate-resistant tumours and twenty-five genes were differentially expressed between castrate-resistant tumours with high and low pARser213 expression.


Combined pARser81 and pARser213 provides a novel prognostic biomarker for castrate-resistant disease and a potential predictive and therapeutic target for prostate cancer. Further studies will be required to investigate the combined effects of targeting AR and PI3K/AKT signalling.

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Fig. 1: Androgen receptor phosphorylation status between hormone-naive and castrate-resistant prostate cancer.
Fig. 2: Combined pARser81 and pARser213 expression in castrate-resistant tumours.
Fig. 3: Stimulating androgen-dependent androgen receptor signalling in LNCaP-AI cells.
Fig. 4: Stimulating androgen-independent androgen receptor signalling in LNCaP-AI cells.
Fig. 5: Inhibiting androgen receptor signalling in LNCaP-AI cells.


  1. 1.

    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin. 2016;66:7–30.

    Article  Google Scholar 

  2. 2.

    Tilki D, Schaeffer EM, Evans CP. Understanding mechanisms of resistance in metastatic castration-resistant prostate cancer: the role of the androgen receptor. Eur Urol Focus. 2016;2:499–505.

    Article  Google Scholar 

  3. 3.

    Ponguta LA, Gregory CW, French FS, Wilson EM. Site-specific androgen receptor serine phosphorylation linked to epidermal growth factor-dependent growth of castration-recurrent prostate cancer. J Biol Chem. 2008;283:20989–1001.

    CAS  Article  Google Scholar 

  4. 4.

    Hsu FN, Chen MC, Chiang MC, Lin E, Lee YT, Huang PH, et al. Regulation of androgen receptor and prostate cancer growth by cyclin-dependent kinase 5. J Biol Chem. 2011;286:33141–9.

    CAS  Article  Google Scholar 

  5. 5.

    Górowska-Wójtowicz E, Hejmej A, Kamińska A, Pardyak L, Kotula-Balak M, Dulińska-Litewka J, et al. Anti-androgen 2-hydroxyflutamide modulates cadherin, catenin and androgen receptor phosphorylation in androgen-sensitive LNCaP and androgen-independent PC3 prostate cancer cell lines acting via PI3K/Akt and MAPK/ERK1/2 pathways. Toxicol Vitr. 2017;40:324–35.

    Article  Google Scholar 

  6. 6.

    McCall P, Gemmell LK, Mukherjee R, Bartlett JM, Edwards J. Phosphorylation of the androgen receptor is associated with reduced survival in hormone-refractory prostate cancer patients. Br J Cancer. 2008;98:1094–101.

    CAS  Article  Google Scholar 

  7. 7.

    Heimberger AB, Hlatky R, Suki D, Yang D, Weinberg J, Gilbert M, et al. Prognostic effect of epidermal growth factor receptor and EGFRvIII in glioblastoma multiforme patients. Clin Cancer Res. 2005;11:1462–6.

    CAS  Article  Google Scholar 

  8. 8.

    Armstrong AJ, Halabi S, Healy P, Alumkal JJ, Winters C, Kephart J, et al. Phase II trial of the PI3 kinase inhibitor buparlisib (BKM-120) with or without enzalutamide in men with metastatic castration resistant prostate cancer. Eur J Cancer. 2017;81:228–36.

    CAS  Article  Google Scholar 

  9. 9.

    Willder JM, Heng SJ, McCall P, Adams CE, Tannahill C, Fyffe G, et al. Androgen receptor phosphorylation at serine 515 by Cdk1 predicts biochemical relapse in prostate cancer patients. Br J Cancer. 2013;108:139–48.

    CAS  Article  Google Scholar 

  10. 10.

    Patek S, Willder J, Heng J, Taylor B, Horgan P, Leung H, et al. Androgen receptor phosphorylation status at serine 578 predicts poor outcome in prostate cancer patients. Oncotarget. 2017;8:4875–87.

    PubMed  Google Scholar 

  11. 11.

    Bartlett JM, Brawley D, Grigor K, Munro AF, Dunne B, Edwards J. Type I receptor tyrosine kinases are associated with hormone escape in prostate cancer. J Pathol. 2005;205:522–9.

    CAS  Article  Google Scholar 

  12. 12.

    Edwards J, Traynor P, Munro AF, Pirret CF, Dunne B, Bartlett JM. The role of HER1-HER4 and EGFRvIII in hormone-refractory prostate cancer. Clin Cancer Res. 2006;12:123–30.

    CAS  Article  Google Scholar 

  13. 13.

    Kirkegaard T, Edwards J, Tovey S, McGlynn LM, Krishna SN, Mukherjee R, et al. Observer variation in immunohistochemical analysis of protein expression, time for a change? Histopathology. 2006;48:787–94.

    CAS  Article  Google Scholar 

  14. 14.

    Trejo CL, Profile VO, Babić M, Imler E, Gonzalez M, Bibikov S, et al. Extraction-free whole transcriptome gene expression analysis of FFPE sections and histology-directed subareas of tissue. bioRxiv. 2018;14:e0212031.

    Google Scholar 

  15. 15.

    Mav D, Shah RR, Howard BE, Auerbach SS, Bushel PR, Collins JB, et al. A hybrid gene selection approach to create the S1500+ targeted gene sets for use in high-throughput transcriptomics. PLoS ONE. 2018;13:e0191105.

    Article  Google Scholar 

  16. 16.

    Thorpe LM, Yuzugullu H, Zhao JJ. PI3K in cancer: divergent roles of isoforms, modes of activation and therapeutic targeting. Nat Rev Cancer. 2015;15:7–24.

    CAS  Article  Google Scholar 

  17. 17.

    Majumder PK, Sellers WR. Akt-regulated pathways in prostate cancer. Oncogene. 2005;24:7465–74.

    CAS  Article  Google Scholar 

  18. 18.

    Lin HK, Wang L, Hu YC, Altuwaijri S, Chang C. Phosphorylation-dependent ubiquitylation and degradation of androgen receptor by Akt require Mdm2 E3 ligase. EMBO J. 2002;21:4037–48.

    CAS  Article  Google Scholar 

  19. 19.

    Wen Y, Hu MC, Makino K, Spohn B, Bartholomeusz G, Yan DH, et al. HER-2/neu promotes androgen-independent survival and growth of prostate cancer cells through the Akt pathway. Cancer Res. 2000;60:6841–5.

    CAS  PubMed  Google Scholar 

  20. 20.

    Blom S, Mäki-Teeri P, Erickson A, Paavolainen L, Mirtti T, Rannikko A, et al. Abstract: PI3K/Akt activity regulates androgen receptor expression and predicts poor clinical outcome in non-metastatic hormone-naïve prostate cancer. Cancer Res. 2017;77:5732–3.

    Google Scholar 

  21. 21.

    Wang Q, Li W, Zhang Y, Yuan X, Xu K, Yu J, et al. Androgen receptor regulates a distinct transcription program in androgen-independent prostate cancer. Cell. 2009;138:245–56.

    CAS  Article  Google Scholar 

  22. 22.

    Yu C, Cao H, He X, Sun P, Feng Y, Chen L, et al. Cyclin-dependent kinase inhibitor 3 (CDKN3) plays a critical role in prostate cancer via regulating cell cycle and DNA replication signaling. Biomed Pharmacother. 2017;96:1109–18.

    CAS  Article  Google Scholar 

  23. 23.

    Zhao H, Whitfield ML, Xu T, Botstein D, Brooks JD. Diverse effects of methylseleninic acid on the transcriptional program of human prostate cancer cells. Mol Biol Cell. 2004;15:506–19.

    CAS  Article  Google Scholar 

  24. 24.

    Maxwell CA, McCarthy J, Turley E. Cell-surface and mitotic-spindle RHAMM: moonlighting or dual oncogenic functions? J Cell Sci. 2008;121:925–32.

    CAS  Article  Google Scholar 

  25. 25.

    Chen S, Xu Y, Yuan X, Bubley GJ, Balk SP. Androgen receptor phosphorylation and stabilization in prostate cancer by cyclin-dependent kinase 1. Proc Natl Acad Sci USA. 2006;103:15969–74.

    CAS  Article  Google Scholar 

  26. 26.

    McClurg UL, Summerscales EE, Harle VJ, Gaughan L, Robson CN. Deubiquitinating enzyme Usp12 regulates the interaction between the androgen receptor and the Akt pathway. Oncotarget. 2014;5:7081–92.

    Article  Google Scholar 

  27. 27.

    Li Y, Xie N, Gleave ME, Rennie PS, Dong X. AR-v7 protein expression is regulated by protein kinase and phosphatase. Oncotarget. 2015;6:33743–54.

    PubMed  PubMed Central  Google Scholar 

  28. 28.

    Dhanasekaran SM, Barrette TR, Ghosh D, Shah R, Varambally S, Kurachi K, et al. Delineation of prognostic biomarkers in prostate cancer. Nature. 2001;412:822–6.

    CAS  Article  Google Scholar 

  29. 29.

    Stamey TA, Warrington JA, Caldwell MC, Chen Z, Fan Z, Mahadevappa M, et al. Molecular genetic profiling of Gleason grade 4/5 prostate cancers compared to benign prostatic hyperplasia. J Urol. 2001;166:2171–7.

    CAS  Article  Google Scholar 

  30. 30.

    Magee JA, Araki T, Patil S, Ehrig T, True L, Humphrey PA, et al. Expression profiling reveals hepsin overexpression in prostate cancer. Cancer Res. 2001;61:5692–6.

    CAS  PubMed  Google Scholar 

  31. 31.

    Chen Z, Fan Z, McNeal JE, Nolley R, Caldwell MC, Mahadevappa M, et al. Hepsin and maspin are inversely expressed in laser capture microdissectioned prostate cancer. J Urol. 2003;169:1316–9.

    CAS  Article  Google Scholar 

  32. 32.

    Craft N, Shostak Y, Carey M, Sawyers CL. A mechanism for hormone-independent prostate cancer through modulation of androgen receptor signaling by the HER-2/neu tyrosine kinase. Nat Med. 1999;5:280–5.

    CAS  Article  Google Scholar 

  33. 33.

    Schroeder RL, Stevens CL, Sridhar J. Small molecule tyrosine kinase inhibitors of ErbB2/HER2/Neu in the treatment of aggressive breast cancer. Molecules. 2014;19:15196–212.

    Article  Google Scholar 

  34. 34.

    Day KC, Lorenzatti Hiles G, Kozminsky M, Dawsey SJ, Paul A, Broses LJ, et al. HER2 and EGFR overexpression support metastatic progression of prostate cancer to bone. Cancer Res. 2017;77:74–85.

    CAS  Article  Google Scholar 

  35. 35.

    Shah RB, Ghosh D, Elder JT. Epidermal growth factor receptor (ErbB1) expression in prostate cancer progression: correlation with androgen independence. Prostate. 2006;66:1437–44.

    CAS  Article  Google Scholar 

  36. 36.

    Traish AM, Morgentaler A. Epidermal growth factor receptor expression escapes androgen regulation in prostate cancer: a potential molecular switch for tumour growth. Br J Cancer. 2009;101:1949–56.

    CAS  Article  Google Scholar 

  37. 37.

    DeHaan AM, Wolters NM, Keller ET, Ignatoski KM. EGFR ligand switch in late stage prostate cancer contributes to changes in cell signaling and bone remodeling. Prostate. 2009;69:528–37.

    CAS  Article  Google Scholar 

  38. 38.

    Narita Y, Nagane M, Mishima K, Huang HJ, Furnari FB, Cavenee WK. Mutant epidermal growth factor receptor signaling down-regulates p27 through activation of the phosphatidylinositol 3-kinase/Akt pathway in glioblastomas. Cancer Res. 2002;62:6764–9.

    CAS  PubMed  Google Scholar 

  39. 39.

    Gottlieb B, Beitel LK, Nadarajah A, Paliouras M, Trifiro M. The androgen receptor gene mutations database: 2012 update. Hum Mutat. 2012;33:887–94.

    CAS  Article  Google Scholar 

  40. 40.

    Hara T, Miyazaki J, Araki H, Yamaoka M, Kanzaki N, Kusaka M, et al. Novel mutations of androgen receptor: a possible mechanism of bicalutamide withdrawal syndrome. Cancer Res. 2003;63:149–53.

    CAS  PubMed  Google Scholar 

  41. 41.

    Korpal M, Korn JM, Gao X, Rakiec DP, Ruddy DA, Doshi S, et al. An F876L mutation in androgen receptor confers genetic and phenotypic resistance to MDV3100 (enzalutamide). Cancer Discov. 2013;3:1030–43.

    CAS  Article  Google Scholar 

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We thank Prostate Cancer UK (S14-003) for funding the research presented. We would like to acknowledge the Glasgow Research Tissue Facility and Glasgow Biorepository for supporting this study.

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Correspondence to Milly J. McAllister.

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McAllister, M.J., McCall, P., Dickson, A. et al. Androgen receptor phosphorylation at serine 81 and serine 213 in castrate-resistant prostate cancer. Prostate Cancer Prostatic Dis (2020).

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