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GEP100 links epidermal growth factor receptor signalling to Arf6 activation to induce breast cancer invasion

Nature Cell Biology volume 10, pages 85–92 (2008)Cite this article

Abstract

Epidermal growth factor (EGF) receptor (EGFR) signalling is implicated in tumour invasion and metastasis1,2. However, whether there are EGFR signalling pathways specifically used for tumour invasion still remains elusive. Overexpression of Arf6 and its effector, AMAP1, correlates with and is crucial for the invasive phenotypes of different breast cancer cells3,4,5,6. Here we identify the mechanism by which Arf6 is activated to induce tumour invasion. We found that GEP100/BRAG2, a guanine nucleotide exchanging factor (GEF) for Arf6, is responsible for the invasive activity of MDA-MB-231 breast cancer cells, whereas the other ArfGEFs are not. GEP100, through its pleckstrin homology domain, bound directly to Tyr1068/1086-phosphorylated EGFR to activate Arf6. Overexpression of GEP100, together with Arf6, caused non-invasive MCF7 cells7 to become invasive, which was dependent on EGF stimulation. Moreover, GEP100 knockdown blocked tumour metastasis. GEP100 was expressed in 70% of primary breast ductal carcinomas, and was preferentially co-expressed with EGFR in the malignant cases. Our results indicate that GEP100 links EGFR signalling to Arf6 activation to induce invasive activities of some breast cancer cells, and hence may contribute to their metastasis and malignancy.

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Figure 1: GEP100 is responsible for the Matrigel invasion activity of MDA-MB-231 cells.
Figure 2: GEP100 associates with ligand-activated EGFR to induce Arf6 activation and tumour invasion.
Figure 3: Co-expression of GEP100 and Arf6 evokes EGF-dependent invasiveness in MCF7 cells.
Figure 4: Silencing of GEP100 blocks lung metastasis in mice.

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Acknowledgements

We thank M. Hiraishi, M. Iwahara and M. Miyoshi for their help, T. Yoneda for 4T1/luc cells, RIKEN BioResource Center, M. Hirata and P. Randazzo for cDNAs, and H. A. Popiel for reading the manuscript. This work was supported in part by grants-in-aid from the Ministry of Education, Science, Sports and Culture of Japan, and by the Mochida Memorial Foundation and the Naito Foundation.

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Author notes

  1. Masaki Morishige and Shigeru Hashimoto: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, Osaka Bioscience Institute, Osaka, 565-0874, Japan

    Masaki Morishige, Shigeru Hashimoto, Shumei Wei, Ari Hashimoto, Atsuko Yamada, Hajime Yano, Yuichi Mazaki & Hisataka Sabe

  2. Department of Neurosurgery, School of Medicine, Oita University, Oita, 879-5593, Japan

    Masaki Morishige & Hidenori Kobayashi

  3. Department of Thoracic Surgery, Faculty of Medicine, Kyoto University, Kyoto, 606-8507, Japan

    Eiji Ogawa & Hiromi Wada

  4. Laboratory of Diagnostic Pathology, Kyoto University Hospital, Kyoto, 606-8501, Japan

    Eiji Ogawa, Hirokazu Kotani & Toshiaki Manabe

  5. Center for Anatomical Studies, Kyoto University Graduate School of Medicine, Kyoto, 606-8507, Japan

    Yoshinobu Toda & Hirokazu Kotani

  6. Laboratory of Supramolecular Crystallography, Institute for Protein Research, Osaka University, Osaka, 565-0871, Japan

    Mayumi Hirose

  7. Kodama Breast Clinic, Kyoto, 603-8325, Japan

    Hiroshi Kodama & Yoshinori Nio

  8. Graduate School of Biosciences, Kyoto University, Kyoto, 606-8607, Japan

    Hisataka Sabe

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Correspondence to Hisataka Sabe.

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Supplementry figures S1, S2, S3, S4, S5, S6 and Methods (PDF 618 kb)

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Morishige, M., Hashimoto, S., Ogawa, E. et al. GEP100 links epidermal growth factor receptor signalling to Arf6 activation to induce breast cancer invasion. Nat Cell Biol 10, 85–92 (2008). https://doi.org/10.1038/ncb1672

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