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Fibronectin induction abrogates the BRAF inhibitor response of BRAF V600E/PTEN-null melanoma cells

Oncogene volume 35, pages 1225–1235 (2016)Cite this article

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Abstract

The mechanisms by which some melanoma cells adapt to Serine/threonine-protein kinase B-Raf (BRAF) inhibitor therapy are incompletely understood. In the present study, we used mass spectrometry-based phosphoproteomics to determine how BRAF inhibition remodeled the signaling network of melanoma cell lines that were BRAF mutant and PTEN null. Short-term BRAF inhibition was associated with marked changes in fibronectin-based adhesion signaling that were PTEN dependent. These effects were recapitulated through BRAF siRNA knockdown and following treatment with chemotherapeutic drugs. Increased fibronectin expression was also observed in mouse xenograft models as well as specimens from melanoma patients undergoing BRAF inhibitor treatment. Analysis of a melanoma tissue microarray showed loss of PTEN expression to predict for a lower overall survival, with a trend for even lower survival being seen when loss of fibronectin was included in the analysis. Mechanistically, the induction of fibronectin limited the responses of these PTEN-null melanoma cell lines to vemurafenib, with enhanced cytotoxicity observed following the knockdown of either fibronectin or its receptor α5β1 integrin. This in turn abrogated the cytotoxic response to BRAF inhibition via increased AKT signaling, which prevented the induction of cell death by maintaining the expression of the pro-survival protein Mcl-1. The protection conveyed by the induction of FN expression could be overcome through combined treatment with a BRAF and PI3K inhibitor.

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Acknowledgements

We thank Gideon Bollag (Plexxikon) for providing vemurafenib and the PLX4720 chow; Noel Clark, Ashley Troutman, Kelli Noyd, Jayme O’Neal and Holly Crandell for assistance with IHC staining; Marta Perez and Monica Torres for assistance with xenograft studies; Agnieszka Kasprazak and Joseph Johnson for assistance with microscopy; and Laura Hall for assistance with qRT–PCR. The work in the Smalley laboratory is supported by R01 CA161107 from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Molecular Oncology, The Moffitt Cancer Center & Research Institute, Tampa, FL, USA

    I V Fedorenko, J M Koomen, E R Wood & K S M Smalley

  2. Department of Cancer Biology, Thomas Jefferson University, Philadelphia, PA, USA

    E V Abel

  3. Department of Proteomics, The Moffitt Cancer Center & Research Institute, Tampa, FL, USA

    B Fang & E R Wood

  4. Department of Biostatistics and Bioinformatics, The Moffitt Cancer Center & Research Institute, Tampa, FL, USA

    Y A Chen & K J Fisher

  5. Department of Cutaneous Oncology, The Moffitt Cancer Center & Research Institute, Tampa, FL, USA

    S Iyengar, V K Sondak, J L Messina, G T Gibney & K S M Smalley

  6. Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, Nashville, TN, USA

    K B Dahlman & J A Sosman

  7. Department of Surgery, Massachusetts General Hospital, Boston, MA, USA

    J A Wargo

  8. Department of Medicine, Massachusetts General Hospital, Boston, MA, USA

    K T Flaherty

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  1. I V Fedorenko
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Fedorenko, I., Abel, E., Koomen, J. et al. Fibronectin induction abrogates the BRAF inhibitor response of BRAF V600E/PTEN-null melanoma cells. Oncogene 35, 1225–1235 (2016). https://doi.org/10.1038/onc.2015.188

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