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

• Published in: Oncogene Vol. 32; no. 34; pp. 3992 - 4000
• Main Authors: Ha, S, Iqbal, N J, Mita, P, Ruoff, R, Gerald, W L, Lepor, H, Taneja, S S, Lee, P, Melamed, J, Garabedian, M J, Logan, S K
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 22.08.2013
• Subjects: Alanine; Amino Acid Substitution; Androgen receptors; Androgens; Antineoplastic Agents, Hormonal - therapeutic use; Blotting, Western; Caveolin; Caveolin 2 - genetics; Caveolin 2 - metabolism; Cell Line, Tumor; Cyclin A1 - genetics; Cyclin A1 - metabolism; DNA microarrays; Drug Resistance, Neoplasm; Gene expression; Gene Expression Regulation, Neoplastic; Genetic aspects; HEK293 Cells; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Hormone receptors; Hormones; Humans; Immunohistochemistry; Insulin-like growth factor-binding protein 5; Interleukin 6; Interleukin-6 - genetics; Interleukin-6 - metabolism; Kinases; Male; Mutants; Mutation; Nkx3.1 protein; Phosphorylation; Physiological aspects; Prostate cancer; Prostate-Specific Antigen - genetics; Prostate-Specific Antigen - metabolism; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism; Prostatic Neoplasms - pathology; Proto-Oncogene Proteins c-pim-1 - genetics; Proto-Oncogene Proteins c-pim-1 - metabolism; Proto-oncogenes; Receptors, Androgen - genetics; Receptors, Androgen - metabolism; Reporter gene; Reverse Transcriptase Polymerase Chain Reaction; Risk factors; Serine; Serine - genetics; Serine - metabolism; Signal transduction; Tissue Array Analysis; Transcription; Transcription Factors - genetics; Transcription Factors - metabolism; Tumor cell lines; United States
• ISSN: 0950-9232; 1476-5594
• DOI: 10.1038/onc.2012.412

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.