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CDCP1 cleavage is necessary for homodimerization-induced migration of triple-negative breast cancer

Oncogene volume 35, pages 4762–4772 (2016)Cite this article

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Abstract

Triple-negative breast cancer (TNBC) is a highly aggressive and metastatic form of breast cancer that lacks the estrogen, progesterone and HER2 receptors and is resistant to targeted and hormone therapies. TNBCs express high levels of the transmembrane glycoprotein, complement C1r/C1s, Uegf, Bmp1 (CUB)-domain containing protein 1 (CDCP1), which has been correlated with the aggressiveness and poor prognosis of multiple carcinomas. Full-length CDCP1 (flCDCP1) can be proteolytically cleaved, resulting in a cleaved membrane-bound isoform (cCDCP1). CDCP1 is phosphorylated by Src family kinases in its full-length and cleaved states, which is important for its pro-metastatic signaling. We observed that cCDCP1, compared with flCDCP1, induced a dramatic increase in phosphorylation of the migration-associated proteins: PKCδ, ERK1/2 and p38 mitogen-activated protein kinase in HEK 293T. In addition, only cCDCP1 induced migration of HEK 293T cells and rescued migration of the TNBC cell lines expressing short hairpin RNA against CDCP1. Importantly, we found that only cCDCP1 is capable of dimerization, which can be blocked by expression of the extracellular portion of cCDCP1 (ECC), indicating that dimerization occurs through CDCP1’s ectodomain. We found that ECC inhibited phosphorylation of PKCδ and migration of TNBC cells in two-dimensional culture. Furthermore, ECC decreased cell invasiveness, inhibited proliferation and stimulated apoptosis of TNBC cells in three-dimensional culture, indicating that the cCDCP1 dimer is an important contributor to TNBC aggressiveness. These studies have important implications for the development of a therapeutic to block CDCP1 activity and TNBC metastasis.

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Acknowledgements

This work was supported by NIH Award F31CA196226 to HJW. We thank Drs Alice Police and Erin Lin for providing primary TNBC tissue, Dr Peter Kaiser for MDA-MB-231 and MDA-MB-468 cell lines, Dr Dan Mercola for MDA-MB-453 cell line, Dr Xing Dai for MCF10A cell line, Dr Albert Koong for split-luciferase vectors, Dr Inder Verma for pVSVG and ΔR8.2 lentivirus packaging plasmids, Dr Jae-Won Soh for pHACE-PKCδ construct, Dr Sanford Shattil for pRC-CMV-Src construct, Dr Peter Chumakov for pLM-CMV lentiviral vector, Miranda Paley from Dr Jennifer Prescher lab and Linan Liu from Dr Weian Zhao’s lab for help with IVIS Lumina Imager, and Dr David Fruman and Dr Hung Fan for critical reading of the manuscript.

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Authors and Affiliations

  1. Department of Molecular Biology and Biochemistry, University of California, Irvine, CA, USA

    H J Wright, M Motazedi, L J Nelson & O V Razorenova

  2. Department of Biomedical Engineering, University of California, Irvine, CA, USA

    J Arulmoli & L A Flanagan

  3. Department of Pathology, Hoag Memorial Hospital Presbyterian, Newport Beach, CA, USA

    F S Heinemann

  4. Department of Neurology, University of California, Irvine, CA, USA

    L A Flanagan

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Wright, H., Arulmoli, J., Motazedi, M. et al. CDCP1 cleavage is necessary for homodimerization-induced migration of triple-negative breast cancer. Oncogene 35, 4762–4772 (2016). https://doi.org/10.1038/onc.2016.7

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