Breast Cancer Research and Treatment

, Volume 131, Issue 2, pp 401–411 | Cite as

Influence of stromal–epithelial interactions on breast cancer in vitro and in vivo

  • Shirley M. Potter
  • Roisin M. Dwyer
  • Marion C. Hartmann
  • Sonja Khan
  • Marie P. Boyle
  • Catherine E. Curran
  • Michael J. Kerin
Preclinical study
First Online:
Received:
Accepted:

Abstract

Stromal cell-secreted chemokines including CCL2 have been implicated in the primary tumor microenvironment, as mediators of tumor cell migration, proliferation, and angiogenesis. Expression of CCL2 and its principal receptor CCR2 was analyzed by RQ-PCR in primary tumor cells and breast cancer cell lines. Breast cancer cell lines (MDA-MB-231, T47D) were co-cultured directly on a monolayer of primary breast tumor and normal stromal cells, retrieved using EpCAM+ magnetic beads, and changes in expression of CCL2, CCR2, MMP11, ELK1, VIL2, and Ki67 detected by RQ-PCR. Epithelial cell migration and proliferation in response to stromal cell-secreted factors was also analyzed. In vivo, tumor xenografts were formed by co-injecting T47D cells with primary tumor stromal cells. Following establishment, tumors were harvested and digested, epithelial cells retrieved and analyzed by RQ-PCR. Whole tumor tissue was also analyzed by immunohistochemistry for CD31 and the VIL2 encoded protein Ezrin. Tumor stromal cells expressed significantly higher levels of CCL2 than normal cells, with no CCR2 expression detected. Primary epithelial cells and breast cancer cell lines expressed elevated CCL2, with relative expression of CCR2 found to be higher than the ligand. Interaction of breast cancer epithelial cells with primary tumor, but not normal stromal cells, stimulated increased expression of CCL2 (8-fold), ELK1 (6-fold), VIL2 (6-fold), and MMP11 (17-fold). Factors secreted by stromal cells, including CCL2, stimulated a significant increase in epithelial cell migration, with no effect on cell proliferation in vitro observed. In vivo, the presence of stromal cells resulted in tumors of increased volume, mediated at least in part through neoangiogenesis demonstrated by immunohistochemistry (CD31). Admixed tumor xenografts exhibited increased expression of Ki67, MMP11, VIL2, and ELK1. Elevated Ezrin protein was also detected, with increased cytoplasmic localization. The results presented highlight mechanisms through which breast cancer epithelial cells can harness stromal cell biology to support tumor progression.

Keywords

Breast cancer Stromal Epithelial Xenograft Angiogenesis Proliferation Metastasis 
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Notes

Acknowledgments

Shirley Potter and Roisin Dwyer were funded by the Health Research Board of Ireland, a Clinical Research Training Fellowship and Research Project Grant, respectively. Marion Hartmann was funded by Breast Cancer Ireland. Funding was also received from the National Breast Cancer Research Institute.

Conflict of interest

The authors declare that they have no competing interests.

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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Shirley M. Potter
    • 1
  • Roisin M. Dwyer
    • 1
  • Marion C. Hartmann
    • 1
  • Sonja Khan
    • 1
  • Marie P. Boyle
    • 1
  • Catherine E. Curran
    • 1
  • Michael J. Kerin
    • 1
  1. 1.Division of Surgery, School of MedicineNational University of IrelandGalwayIreland

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