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Dasatinib inhibits site-specific tyrosine phosphorylation of androgen receptor by Ack1 and Src kinases

Oncogene volume 29, pages 3208–3216 (2010)Cite this article

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Abstract

Activation of androgen receptor (AR) may have a role in the development of castration-resistant prostate cancer. Two intracellular tyrosine kinases, Ack1 (activated cdc42-associated kinase) and Src, phosphorylate and enhance AR activity and promote prostate xenograft tumor growth in castrated animals. However, the upstream signals that activate these kinases and lead to AR activation are incompletely characterized. In this study, we investigated AR phosphorylation in response to non-androgen ligand stimulation using phospho-specific antibodies. Treatment of LNCaP and LAPC-4 cells with epidermal growth factor (EGF), heregulin, Gas6 (ligand binding to the Mer receptor tyrosine kinase and activating Ack1 downstream), interleukin (IL)-6 or bombesin stimulated cell proliferation in the absence of androgen. Treatment of LNCaP and LAPC-4 cells with EGF, heregulin or Gas6 induced AR phosphorylation at Tyr-267, whereas IL-6 or bombesin treatment did not. AR phosphorylation at Tyr-534 was induced by treatment with EGF, IL-6 or bombesin, but not by heregulin or Gas6. Small interfering RNA-mediated knockdown of Ack1 or Src showed that Ack1 mediates heregulin- and Gas6-induced AR Tyr-267 phosphorylation, whereas Src mediates Tyr-534 phosphorylation induced by EGF, IL-6 and bombesin. Dasatinib, a Src inhibitor, blocked EGF-induced Tyr-534 phosphorylation. In addition, we showed that dasatinib also inhibited Ack1 kinase. Dasatinib inhibited heregulin-induced Ack1 kinase activity and AR Tyr-267 phosphorylation. In addition, dasatinib inhibited heregulin-induced AR-dependent reporter activity. Dasatinib also inhibited heregulin-induced expression of endogenous AR target genes. Dasatinib inhibited Ack1-dependent colony formation and prostate xenograft tumor growth in castrated mice. Interestingly, Ack1 or Src knockdown or dasatinib did not inhibit EGF-induced AR Tyr-267 phosphorylation or EGF-stimulated AR activity, suggesting the existence of an additional tyrosine kinase that phosphorylates AR at Tyr-267. These data suggest that specific tyrosine kinases phosphorylate AR at distinct sites and that dasatinib may exert antitumor activity in prostate cancer through inhibition of Ack1.

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Acknowledgements

We thank Francis Lee of Bristol-Myers-Squibb for providing dasatinib, Charlene Ross and staff of the UNC Lineberger Animal Studies Facility for assistance in xenograft tumor experiments, Dominic Moore for statistical analysis and Nupam Mahajan for discussions and sharing unpublished papers. This study was supported by grants from NIH R01CA120921 (YEW), NIH R01CA120304 (HSE) and NIH T32ES007017 (MK).

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Authors and Affiliations

  1. UNC Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Y Liu, Z Zhang, H S Earp & Y E Whang

  2. Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    M Karaca & Y E Whang

  3. Department of Microbiology, University of Virginia Health System, Charlottesville, VA, USA

    D Gioeli

  4. Department of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    H S Earp & Y E Whang

  5. Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    H S Earp

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  1. Y Liu
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Correspondence to Y E Whang.

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Liu, Y., Karaca, M., Zhang, Z. et al. Dasatinib inhibits site-specific tyrosine phosphorylation of androgen receptor by Ack1 and Src kinases. Oncogene 29, 3208–3216 (2010). https://doi.org/10.1038/onc.2010.103

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