The Transmodulation of HER2 and EGFR by Substance P in Breast Cancer Cells Requires c-Src and Metalloproteinase Activation

PLoS One. 2015 Jun 26;10(6):e0129661. doi: 10.1371/journal.pone.0129661. eCollection 2015.

Abstract

Background: Substance P (SP) is a pleiotropic cytokine/neuropeptide that enhances breast cancer (BC) aggressiveness by transactivating tyrosine kinase receptors like EGFR and HER2. We previously showed that SP and its cognate receptor NK-1 (SP/NK1-R) signaling modulates the basal phosphorylation of HER2 and EGFR in BC, increasing aggressiveness and drug resistance. In order to elucidate the mechanisms responsible for NK-1R-mediated HER2 and EGFR transactivation, we investigated the involvement of c-Src (a ligand-independent mediator) and of metalloproteinases (ligand-dependent mediators) in HER2/EGFR activation.

Results and discussion: Overexpression of NK-1R in MDA-MB-231 and its chemical inhibition in SK-BR-3, BT-474 and MDA-MB-468 BC cells significantly modulated c-Src activation, suggesting that this protein is a mediator of NK-1R signaling. In addition, the c-Src inhibitor 4-(4'-phenoxyanilino)-6,7-dimethoxyquinazoline prevented SP-induced activation of HER2. On the other hand, SP-dependent phosphorylation of HER2 and EGFR decreased substantially in the presence of the MMP inhibitor 1-10, phenanthroline monohydrate, and the dual inhibition of both c-Src and MMP almost abolished the activation of HER2 and EGFR. Moreover, the use of these inhibitors demonstrated that this Src and MMP-dependent signaling is important to the cell viability and migration capacity of HER2+ and EGFR+ cell lines.

Conclusion: Our results indicate that the transactivation of HER2 and EGFR by the pro-inflammatory cytokine/neuropeptide SP in BC cells is a c-Src and MMP-dependent process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / genetics
  • Breast Neoplasms / metabolism*
  • CSK Tyrosine-Protein Kinase
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Survival / drug effects
  • Enzyme Activation
  • ErbB Receptors / metabolism*
  • Female
  • Gene Expression
  • Humans
  • Metalloproteases / antagonists & inhibitors
  • Metalloproteases / metabolism*
  • Neurokinin-1 Receptor Antagonists / pharmacology
  • Phosphorylation
  • Protein Binding
  • Receptor, ErbB-2 / metabolism*
  • Receptors, Neurokinin-1 / genetics
  • Receptors, Neurokinin-1 / metabolism
  • Substance P / metabolism*
  • Trans-Activators / metabolism
  • src-Family Kinases / antagonists & inhibitors
  • src-Family Kinases / metabolism*

Substances

  • Neurokinin-1 Receptor Antagonists
  • Receptors, Neurokinin-1
  • Trans-Activators
  • Substance P
  • ErbB Receptors
  • Receptor, ErbB-2
  • CSK Tyrosine-Protein Kinase
  • src-Family Kinases
  • CSK protein, human
  • Metalloproteases

Grants and funding

This work was supported by a grant from the Fondo de Investigación Sanitaria (PI08022) (P.G.), Instituto de salud Carlos III-Subdireción General de Evaluación y Fomento de Investigación, Fondo Europeo de Desarollo Regional, Unión Europea, Una manera de hacer Europa (http://www.eng.isciii.es/), and by a grant from the Fundación Cellex (P.G), by Redes Temáticas de Investigación en Cáncer (RTICC, RD07/0020/2014) (P.G) (http://www.rticc.org/). The work was carried out in part at the Esther Koplowitz Centre, Barcelona. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.