Get access
Journal of Cellular Biochemistry

Stem cell property epithelial-to-mesenchymal transition is a core transcriptional network for predicting cetuximab (Erbitux™) efficacy in KRAS wild-type tumor cells

Authors

  • Cristina Oliveras-Ferraros,

    1. Unit of Translational Research, Catalan Institute of Oncology (ICO), Girona, Spain
    2. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    Search for more papers by this author
  • Alejandro Vazquez-Martin,

    1. Unit of Translational Research, Catalan Institute of Oncology (ICO), Girona, Spain
    2. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    Search for more papers by this author
  • Sílvia Cufí,

    1. Unit of Translational Research, Catalan Institute of Oncology (ICO), Girona, Spain
    2. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    Search for more papers by this author
  • Bernardo Queralt,

    1. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    2. Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Spain
    Search for more papers by this author
  • Luciana Báez,

    1. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    2. Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Spain
    Search for more papers by this author
  • Raquel Guardeño,

    1. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    2. Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Spain
    Search for more papers by this author
  • Xavier Hernández-Yagüe,

    1. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    2. Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Spain
    Search for more papers by this author
  • Begoña Martin-Castillo,

    1. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    2. Unit of Clinical Research, Catalan Institute of Oncology (ICO), Girona, Spain
    Search for more papers by this author
  • Joan Brunet,

    1. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    2. Medical Oncology, Catalan Institute of Oncology (ICO), Girona, Spain
    Search for more papers by this author
  • Javier A. Menendez

    Corresponding author
    1. Unit of Translational Research, Catalan Institute of Oncology (ICO), Girona, Spain
    2. Girona Biomedical Research Institute (IdIBGi), Girona, Spain
    • Catalan Institute of Oncology, Girona (ICO-Girona), Dr. Josep Trueta University Hospital, Avenida de França s/n, E-17007 Girona, Catalonia, Spain.
    Search for more papers by this author

  • Conflicts of Interest: Cristina Oliveras-Ferraros received a research salary from a Grant Award by the “Fundacion Salud 2000,” which is promoted by Merck Serono (Madrid, Spain). All other authors: None to declare.

Abstract

Beyond a well-recognized effect of KRAS mutations in determining de novo inefficacy of cetuximab (CTX) in metastatic colorectal cancer, we urgently need a biomarker signature for predicting CTX efficacy in KRAS wild-type (WT) tumors. CTX-adapted EGFR gene-amplified KRAS WT tumor cell populations were induced by stepwise-chronic exposure of A431 epidermoid cancer cells to CTX. Genome-wide analyses of 44K Agilent's whole human arrays were bioinformatically evaluated by Gene Set Enrichment Analysis (GSEA)-based screening of the KEGG pathway database. Molecular functioning of CTX was found to depend on: (i) The occurrence of a positive feedback loop on Epidermal Growth Factor Receptor (EGFR) activation driven by genes coding for EGFR ligands (e.g., amphiregulin); (ii) the lack of a negative feedback on mitogen-activated protein kinase (MAPK) activation regulated by dual-specificity phosphatases (e.g., DUSP6) and; (iii) the transcriptional status of gene pathways controlling the epithelial-to-mesenchymal transition (EMT) and its reversal (MET) program (actin cytoskeleton and cell–cell communication—e.g., keratins—focal adhesion signaling—e.g., integrins—and EMT-inducing cytokines – e.g., transforming growth factor-β). Quantitative real-time PCR, high-content immunostaining, and flow-cytometry analyses confirmed that CTX efficacy depends on its ability to promote: (i) Stronger cell–cell contacts by up-regulating the expression of the epithelial markers E-cadherin and occludin; (ii) down-regulation of the epithelial transcriptional repressors Zeb, Snail, and Slug accompanied by restoration of cortical F-actin; and (iii) complete prevention of the CD44pos/CD24neg/low mesenchymal immunophenotype. The impact of EGFR ligands/MAPK phosphatases gene transcripts in predicting CTX efficacy in KRAS WT tumors may be tightly linked with the ability of CTX to concurrently reverse the EMT status, a pivotal property of migrating cancer stem cells. J. Cell. Biochem. 112: 10–29, 2011. © 2010 Wiley-Liss, Inc.

Get access to the full text of this article

Ancillary