Cdgap is required for transforming growth factor β- and neu/erbb-2-induced breast cancer cell motility and invasion

ABSTRACT RhoA, Rac1 and Cdc42, the best-characterized members of the Rho family of small GTPases, are critical regulators of many cellular activities. Cdc42 GTPase-activating protein (CdGAP)

is a serine- and proline-rich RhoGAP protein showing GAP activity against both Cdc42 and Rac1 but not RhoA. CdGAP is phosphorylated downstream of the MEK–ERK (extracellular signal-regulated

kinase) pathway in response to serum and is required for normal cell spreading and polarized lamellipodia formation. In this study, we found that CdGAP protein and mRNA levels are highly

increased in mammary tumor explants expressing an activated Neu/ErbB-2 (Neu-NT) receptor. In response to transforming growth factor-β (TGFβ) stimulation, Neu-NT-expressing mammary tumor

explants demonstrate a clear induction in cell motility and invasion. We show that downregulation of CdGAP expression by small interfering RNA abrogates the ability of TGFβ to induce cell

motility and invasion of Neu-NT-expressing mammary tumor explants. However, it has no effect on TGFβ-mediated cell adhesion on type 1 collagen and fibronectin. Interestingly, protein

expression of E-Cadherin is highly increased in Neu-NT-expressing mammary tumor explants depleted of CdGAP. In addition, complete loss of E-Cadherin expression is not observed in

CdGAP-depleted cells during TGFβ-mediated epithelial to mesenchymal transition. Downregulation of the CdGAP expression also decreases cell proliferation of Neu-NT-expressing mammary tumor

explants independently of TGFβ. Rescue analysis using re-expression of various CdGAP deletion-mutant proteins revealed that the proline-rich domain (PRD) but not the GAP domain of CdGAP is

essential to mediate TGFβ-induced cell motility and invasion. Finally, we found that TGFβ induces the expression and phosphorylation of CdGAP in mammary epithelial NMuMG cells. Taken

together, these studies identify CdGAP as a novel molecular target in TGFβ signaling and implicate CdGAP as an essential component in the synergistic interaction between TGFβ and Neu/ErbB-2

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adverse side effects. _J Clin Invest_ 109: 1607–1615. Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS This research was supported by grants from Canadian Institute of

Health Research (CIHR) MOP-84449 to NL-V and the Cancer Research Society to PMS. JJN is supported by a studentship from the Research Institute of the McGill University Health Centre. MP was

a recipient of a CIHR Canada Graduate scholarship. NL-V is a recipient of a FRSQ chercheur-boursier senior. PMS is a research scientist of the Canadian Cancer Society. RDM is a British Heart

Foundation Intermediate Research Fellow (BHF-FS/07/036). AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada Y

He, M Primeau & N Lamarche-Vane * Department of Biochemistry, McGill University, Montreal, Quebec, Canada J J Northey & P M Siegel * Goodman Cancer Research Centre, McGill

University, Montreal, Quebec, Canada J J Northey & P M Siegel * Department of Medical and Molecular Genetics, King's College London, School of Medicine, London, UK R D Machado &

 R Trembath * Department of Clinical Genetics, Guy's Hospital, London, UK R Trembath Authors * Y He View author publications You can also search for this author inPubMed Google Scholar

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PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE He, Y., Northey, J., Primeau, M. _et al._ CdGAP is required for transforming growth factor β- and Neu/ErbB-2-induced

breast cancer cell motility and invasion. _Oncogene_ 30, 1032–1045 (2011). https://doi.org/10.1038/onc.2010.477 Download citation * Received: 06 April 2010 * Revised: 12 August 2010 *

Accepted: 08 September 2010 * Published: 01 November 2010 * Issue Date: 03 March 2011 * DOI: https://doi.org/10.1038/onc.2010.477 SHARE THIS ARTICLE Anyone you share the following link with

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content-sharing initiative KEYWORDS * Rho GTPases * RhoGAPs * TGFβ * breast cancer * cell migration and invasion