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Gab2 and Src co-operate in human mammary epithelial cells to promote growth factor independence and disruption of acinar morphogenesis

Oncogene volume 27, pages 2693–2704 (2008)Cite this article

Abstract

The Gab2 docking protein is a target of several oncogenic protein tyrosine kinases and potentiates activation of the Ras/extracellular signal regulated kinase and phosphatidylinositol 3-kinase (PI3-kinase) pathways. Since Gab2 is phosphorylated by c-Src, and both proteins are overexpressed in breast cancers, we have determined the biological consequences of their co-expression in the immortalized human mammary epithelial cell line MCF-10A. While overexpression of c-Src did not affect acinar morphogenesis or growth factor dependence in three-dimensional culture, c-Src co-operated with Gab2 to promote epidermal growth factor (EGF)-independent acinar growth. In contrast, expression of v-Src or the activated mutant c-SrcY527F led to a spectrum of aberrant phenotypes ranging from spheroids with incomplete luminal clearance to highly disrupted, dispersed structures. Gab2 co-expression shifted the phenotypic distribution towards the dispersed phenotype, an effect not observed with a Gab2 mutant unable to bind the p85 subunit of PI3-kinase (Gab2Δp85). In v-Src-expressing cells, Gab2, but not Gab2Δp85, significantly decreased E-cadherin adhesive strength without altering its surface expression. Gab2 associated with E-cadherin in the presence and absence of v-Src, indicating that the ability of Gab2 to weaken the strength of cell–cell contacts may reflect enhanced activation of PI3-kinase at adherens junctions. Gab2 also increased migration and invasion of these cells in transwell assays, but these effects were p85-independent. Overall, these findings demonstrate a novel mechanism whereby Gab2 may promote metastatic spread and indicate that Gab2 may play several roles during breast cancer progression.

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Acknowledgements

This work was supported by Department of Defense Breast Cancer Research Program Grant DAMD17-00-1-0251 and the National Health and Medical Research Council of Australia. HLB and AJ are the recipients of a Australian Postgraduate Award and NHMRC Dora Lush Biomedical Postgraduate Research Scholarship, respectively.

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Authors and Affiliations

  1. Cancer Research Program, Garvan Institute of Medical Research, Sydney, NSW, Australia

    H L Bennett, T Brummer & R J Daly

  2. Institute for Molecular Bioscience, Queensland Bioscience Precinct, University of Queensland, Queensland, Australia

    A Jeanes & A S Yap

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  1. H L Bennett
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Correspondence to R J Daly.

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Bennett, H., Brummer, T., Jeanes, A. et al. Gab2 and Src co-operate in human mammary epithelial cells to promote growth factor independence and disruption of acinar morphogenesis. Oncogene 27, 2693–2704 (2008). https://doi.org/10.1038/sj.onc.1210928

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