Paracrine recruitment and activation of fibroblasts by c-Myc expressing breast epithelial cells through the IGFs/IGF-1R axis

Anna De Vincenzo, Stefania Belli, Paola Franco, Marialucia Telesca, Ingram Iaccarino, Gerardo Botti, Maria V. Carriero, Marie Ranson, Maria Patrizia Stoppelli

Research output: Contribution to journalArticle

Abstract

Fibroblasts are among the most abundant stromal cells in the tumor microenvironment (TME), progressively differentiating into activated, motile, myofibroblast-like, protumorigenic cells referred to as Cancer-Associated Fibroblasts (CAFs). To investigate the mechanisms by which epithelial cells direct this transition, the early stages of tumorigenesis were exemplified by indirect cocultures of WI-38 or human primary breast cancer fibroblasts with human mammary epithelial cells expressing an inducible c-Myc oncogene (MCF10A-MycER). After c-Myc activation, the conditioned medium (CM) of MCF10A-MycER cells significantly enhanced fibroblast activation and mobilization. As this was accompanied by decreased insulin-like growth factor binding protein-6 (IGFBP-6) and increased insulin-like growth factor-1 and IGF-II (IGF-I, IGF-II) in the CM, IGFs were investigated as key chemotactic factors. Silencing IGFBP-6 or IGF-I or IGF-II expression in epithelial cells or blocking Insulin-like growth factor 1 receptor (IGF-1R) activity on fibroblasts significantly altered fibroblast mobilization. Exposure of WI-38 fibroblasts to CM from induced MCF10A-MycER cells or to IGF-II upregulated FAK phosphorylation on Tyr397, as well as the expression of α-smooth muscle actin (α-SMA), features associated with CAF phenotype and increased cell migratory/invasive behavior. In three-dimensional (3D)-organotypic assays, WI-38 or human primary fibroblasts, preactivated with either CM from MCF10A-MycER cells or IGFs, resulted in a permissive TME that enabled nontransformed MCF10A matrix invasion. This effect was abolished by inhibiting IGF-1R activity. Thus, breast epithelial cell oncogenic activation and stromal fibroblast transition to CAFs are linked through the IGFs/IGF-1R axis, which directly promotes TME remodeling and increases tumor invasion.

Original languageEnglish
JournalInternational Journal of Cancer
DOIs
Publication statusPublished - Jan 1 2019

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Somatomedin Receptors
Breast
Fibroblasts
Epithelial Cells
Insulin-Like Growth Factor II
Conditioned Culture Medium
Tumor Microenvironment
Insulin-Like Growth Factor Binding Protein 6
Insulin-Like Growth Factor I
myc Genes
Myofibroblasts
Chemotactic Factors
Somatomedins
Stromal Cells
Coculture Techniques
Smooth Muscle
Actins
Carcinogenesis
Phosphorylation
Breast Neoplasms

Keywords

  • breast cancer cell invasion
  • c-Myc
  • cancer-associated fibroblasts
  • IGFs/IGF-1R axis
  • Urokinase receptor

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Paracrine recruitment and activation of fibroblasts by c-Myc expressing breast epithelial cells through the IGFs/IGF-1R axis. / De Vincenzo, Anna; Belli, Stefania; Franco, Paola; Telesca, Marialucia; Iaccarino, Ingram; Botti, Gerardo; Carriero, Maria V.; Ranson, Marie; Stoppelli, Maria Patrizia.

In: International Journal of Cancer, 01.01.2019.

Research output: Contribution to journalArticle

De Vincenzo, Anna ; Belli, Stefania ; Franco, Paola ; Telesca, Marialucia ; Iaccarino, Ingram ; Botti, Gerardo ; Carriero, Maria V. ; Ranson, Marie ; Stoppelli, Maria Patrizia. / Paracrine recruitment and activation of fibroblasts by c-Myc expressing breast epithelial cells through the IGFs/IGF-1R axis. In: International Journal of Cancer. 2019.
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AU - De Vincenzo, Anna

AU - Belli, Stefania

AU - Franco, Paola

AU - Telesca, Marialucia

AU - Iaccarino, Ingram

AU - Botti, Gerardo

AU - Carriero, Maria V.

AU - Ranson, Marie

AU - Stoppelli, Maria Patrizia

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AB - Fibroblasts are among the most abundant stromal cells in the tumor microenvironment (TME), progressively differentiating into activated, motile, myofibroblast-like, protumorigenic cells referred to as Cancer-Associated Fibroblasts (CAFs). To investigate the mechanisms by which epithelial cells direct this transition, the early stages of tumorigenesis were exemplified by indirect cocultures of WI-38 or human primary breast cancer fibroblasts with human mammary epithelial cells expressing an inducible c-Myc oncogene (MCF10A-MycER). After c-Myc activation, the conditioned medium (CM) of MCF10A-MycER cells significantly enhanced fibroblast activation and mobilization. As this was accompanied by decreased insulin-like growth factor binding protein-6 (IGFBP-6) and increased insulin-like growth factor-1 and IGF-II (IGF-I, IGF-II) in the CM, IGFs were investigated as key chemotactic factors. Silencing IGFBP-6 or IGF-I or IGF-II expression in epithelial cells or blocking Insulin-like growth factor 1 receptor (IGF-1R) activity on fibroblasts significantly altered fibroblast mobilization. Exposure of WI-38 fibroblasts to CM from induced MCF10A-MycER cells or to IGF-II upregulated FAK phosphorylation on Tyr397, as well as the expression of α-smooth muscle actin (α-SMA), features associated with CAF phenotype and increased cell migratory/invasive behavior. In three-dimensional (3D)-organotypic assays, WI-38 or human primary fibroblasts, preactivated with either CM from MCF10A-MycER cells or IGFs, resulted in a permissive TME that enabled nontransformed MCF10A matrix invasion. This effect was abolished by inhibiting IGF-1R activity. Thus, breast epithelial cell oncogenic activation and stromal fibroblast transition to CAFs are linked through the IGFs/IGF-1R axis, which directly promotes TME remodeling and increases tumor invasion.

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KW - Urokinase receptor

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