Loss of transforming growth factor-beta signaling in mammary fibroblasts enhances CCL2 secretion to promote mammary tumor progression through macrophage-dependent and -independent mechanisms

Neoplasia. 2010 May;12(5):425-33. doi: 10.1593/neo.10200.

Abstract

Whereas the accumulation of fibroblasts and macrophages in breast cancer is a well-documented phenomenon and correlates with metastatic disease, the functional contributions of these stromal cells on breast cancer progression still remain largely unclear. Previous studies have uncovered a potentially important role for CCL2 inflammatory chemokine signaling in regulating metastatic disease through a macrophage-dependent mechanism. In these studies, we demonstrate a significant regulatory mechanism for CCL2 expression in fibroblasts in mediating mammary tumor progression and characterize multiple functions for CCL2 in regulating stromal-epithelial interactions. Targeted ablation of the transforming growth factor-beta (TGF-beta) type 2 receptor in fibroblasts (Tgfbr2(FspKO)) results in a high level of secretion of CCL2, and cografts of Tgfbr2(FspKO) fibroblasts with 4T1 mammary carcinoma cells enhanced tumor progression associated with recruitment of tumor-associated macrophages (TAMs). Antibody neutralization of CCL2 in tumor-bearing mice inhibits primary tumor growth and liver metastases as evidenced by reduced cell proliferation, survival, and TAM recruitment. Both high and low stable expressions of small interfering RNA to CCL2 in Tgfbr2(FspKO) fibroblasts significantly reduce liver metastases without significantly affecting primary tumor growth, cell proliferation, or TAM recruitment. High but not low knockdown of CCL2 enhances tumor cell apoptosis. These data indicate that CCL2 enhances primary tumor growth, survival, and metastases in a dose-dependent manner, through TAM-dependent and -independent mechanisms, with important implications on the potential effects of targeting CCL2 chemokine signaling in the metastatic disease.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Separation
  • Chemokine CCL2 / metabolism*
  • Coculture Techniques
  • Disease Progression
  • Female
  • Fibroblasts / immunology
  • Fibroblasts / metabolism*
  • Flow Cytometry
  • Gene Knockout Techniques
  • Immunohistochemistry
  • Macrophages / metabolism
  • Mammary Glands, Animal / immunology
  • Mammary Glands, Animal / metabolism
  • Mammary Glands, Animal / pathology
  • Mammary Neoplasms, Experimental / immunology
  • Mammary Neoplasms, Experimental / metabolism*
  • Mammary Neoplasms, Experimental / pathology
  • Mice
  • Mice, Nude
  • Neoplasm Metastasis / immunology
  • Neoplasm Metastasis / pathology
  • RNA, Small Interfering
  • Signal Transduction / physiology*
  • Transforming Growth Factor beta / metabolism*

Substances

  • Chemokine CCL2
  • RNA, Small Interfering
  • Transforming Growth Factor beta