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PKCι regulates nuclear YAP1 localization and ovarian cancer tumorigenesis

Oncogene volume 36, pages 534–545 (2017)Cite this article

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Abstract

Atypical protein kinase Cι (PKCι) is an oncogene in lung and ovarian cancer. The PKCι gene PRKCI is targeted for frequent tumor-specific copy number gain (CNG) in both lung squamous cell carcinoma (LSCC) and ovarian serous carcinoma (OSC). We recently demonstrated that in LSCC cells PRKCI CNG functions to drive transformed growth and tumorigenicity by activating PKCι-dependent cell autonomous Hedgehog (Hh) signaling. Here, we assessed whether OSC cells harboring PRKCI CNG exhibit similar PKCι-dependent Hh signaling. Surprisingly, we find that whereas PKCι is required for the transformed growth of OSC cells harboring PRKCI CNG, these cells do not exhibit PKCι-dependent Hh signaling or Hh-dependent proliferation. Rather, transformed growth of OSC cells is regulated by PKCι-dependent nuclear localization of the oncogenic transcription factor, YAP1. Lentiviral shRNA-mediated knockdown (KD) of PKCι leads to decreased nuclear YAP1 and increased YAP1 binding to angiomotin (AMOT), which sequesters YAP1 in the cytoplasm. Biochemical analysis reveals that PKCι directly phosphorylates AMOT at a unique site, Thr750, whose phosphorylation inhibits YAP1 binding. Pharmacologic inhibition of PKCι decreases YAP1 nuclear localization and blocks OSC tumor growth in vitro and in vivo. Immunohistochemical analysis reveals a strong positive correlation between tumor PKCι expression and nuclear YAP1 in primary OSC tumor samples, indicating the clinical relevance of PKCι–YAP1 signaling. Our results uncover a novel PKCι–AMOT–YAP1 signaling axis that promotes OSC tumor growth, and provide a rationale for therapeutic targeting of this pathway for treatment of OSC.

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Acknowledgements

We thank Dr Kim Kalli and the Mayo Clinic Ovarian SPORE Tissue shared resource for ovarian cancer tissue microarrays, The Mayo Clinic Cytogenetics Core for FISH analysis, Ms Brandy Edenfield for tissue processing and immunohistochemical staining, and Kayla Lewis, Capella Weems and Dr Christelle Colin Cassin for technical assistance. This research was supported by the Mayo Clinic Specialized Program in Research Excellence (SPORE) grant CA136393 from the National Institutes of Health (APF is a Project Leader on this grant), a post-doctoral fellowship award from the Ovarian Cancer Research Fund to YW, National Cancer Institute grants R21 CA204938-01 (to VJ) and R01 CA081436-18 (to APF), and the George Haub Family Career Development Award (to VJ). APF is the Monica Flynn Jacoby Professor of Cancer Research, an endowment fund that provides partial support for his research program.

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  1. Y Wang and V Justilien: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cancer Biology, Mayo Clinic College of Medicine, Jacksonville, FL, USA

    Y Wang, V Justilien, K I Brennan, L Jamieson, N R Murray & A P Fields

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Correspondence to N R Murray or A P Fields.

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Wang, Y., Justilien, V., Brennan, K. et al. PKCι regulates nuclear YAP1 localization and ovarian cancer tumorigenesis. Oncogene 36, 534–545 (2017). https://doi.org/10.1038/onc.2016.224

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