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Phosphorylation of the androgen receptor by PIM1 in hormone refractory prostate cancer

Oncogene volume 32, pages 3992–4000 (2013)Cite this article

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Abstract

Integration of cellular signaling pathways with androgen receptor (AR) signaling can be achieved through phosphorylation of AR by cellular kinases. However, the kinases responsible for phosphorylating the AR at numerous sites and the functional consequences of AR phosphorylation are only partially understood. Bioinformatic analysis revealed AR serine 213 (S213) as a putative substrate for PIM1, a kinase overexpressed in prostate cancer. Therefore, phosphorylation of AR serine 213 by PIM1 was examined using a phosphorylation site-specific antibody. Wild-type PIM1, but not catalytically inactive PIM1, specifically phosphorylated AR but not an AR serine-to-alanine mutant (S213A). In vitro kinase assays confirmed that PIM1 can phosphorylate AR S213 in a ligand-independent manner and cell type-specific phosphorylation was observed in prostate cancer cell lines. Upon PIM1 overexpression, AR phosphorylation was observed in the absence of hormone and was further increased in the presence of hormone in LNCaP, LNCaP-abl and VCaP cells. Moreover, phosphorylation of AR was reduced in the presence of PIM kinase inhibitors. An examination of AR-mediated transcription showed that reporter gene activity was reduced in the presence of PIM1 and wild-type AR, but not S213A mutant AR. Androgen-mediated transcription of endogenous PSA, Nkx3.1 and IGFBP5 was also decreased in the presence of PIM1, whereas IL6, cyclin A1 and caveolin 2 were increased. Immunohistochemical analysis of prostate cancer tissue microarrays showed significant P-AR S213 expression that was associated with hormone refractory prostate cancers, likely identifying cells with catalytically active PIM1. In addition, prostate cancers expressing a high level of P-AR S213 were twice as likely to be from biochemically recurrent cancers. Thus, AR phosphorylation by PIM1 at S213 impacts gene transcription and is highly prevalent in aggressive prostate cancer.

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Acknowledgements

We acknowledge funding from the New York University School of Medicine Department of Urology at New York University School of Medicine. This work was supported by NIH R01CA112226 (SL), funds from the NYU Department of Urology, DOD grant PC094786 for the Prostate Cancer Biorepository Network (PCBN, JM), DOD grant PC080010 (PL) and NIH UO1 1U01CA149556-01(PL).

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Authors and Affiliations

  1. Department of Urology, New York University School of Medicine, New York, NY, USA

    S Ha, N J Iqbal, P Mita, R Ruoff, H Lepor, S S Taneja, M J Garabedian & S K Logan

  2. Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, NY, USA

    S Ha, R Ruoff & S K Logan

  3. Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY, USA

    W L Gerald

  4. Department of Pathology, New York University School of Medicine, New York, NY, USA

    P Lee & J Melamed

  5. Department of Microbiology, New York University School of Medicine, New York, NY, USA

    M J Garabedian

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Ha, S., Iqbal, N., Mita, P. et al. Phosphorylation of the androgen receptor by PIM1 in hormone refractory prostate cancer. Oncogene 32, 3992–4000 (2013). https://doi.org/10.1038/onc.2012.412

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