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Cross-talk between endothelial and breast cancer cells regulates reciprocal expression of angiogenic factors in vitro.

Publication ,  Journal Article
Buchanan, CF; Szot, CS; Wilson, TD; Akman, S; Metheny-Barlow, LJ; Robertson, JL; Freeman, JW; Rylander, MN
Published in: J Cell Biochem
April 2012

Reciprocal growth factor exchange between endothelial and malignant cells within the tumor microenvironment may directly stimulate neovascularization; however, the role of host vasculature in regulating tumor cell activity is not well understood. While previous studies have examined the angiogenic response of endothelial cells to tumor-secreted factors, few have explored tumor response to endothelial cells. Using an in vitro co-culture system, we investigated the influence of endothelial cells on the angiogenic phenotype of breast cancer cells. Specifically, VEGF, ANG1, and ANG2 gene and protein expression were assessed. When co-cultured with microvascular endothelial cells (HMEC-1), breast cancer cells (MDA-MB-231) significantly increased expression of ANG2 mRNA (20-fold relative to MDA-MB-231 monoculture). Moreover, MDA-MB-231/HMEC-1 co-cultures produced significantly increased levels of ANG2 (up to 580 pg/ml) and VEGF protein (up to 38,400 pg/ml) while ANG1 protein expression was decreased relative to MDA-MB-231 monocultures. Thus, the ratio of ANG1:ANG2 protein, a critical indicator of neovascularization, shifted in favor of ANG2, a phenomenon known to correlate with vessel destabilization and sprouting in vivo. This angiogenic response was not observed in nonmalignant breast epithelial cells (MCF-10A), where absolute protein levels of MCF-10A/HMEC-1 co-cultures were an order of magnitude less than that of the MDA-MB-231/HMEC-1 co-cultures. Results were further verified with a functional angiogenesis assay demonstrating well-defined microvascular endothelial cell (TIME) tube formation when cultured in media collected from MDA-MB-231/HMEC-1 co-cultures. This study demonstrates that the angiogenic activity of malignant mammary epithelial cells is significantly enhanced by the presence of endothelial cells.

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Published In

J Cell Biochem

DOI

EISSN

1097-4644

Publication Date

April 2012

Volume

113

Issue

4

Start / End Page

1142 / 1151

Location

United States

Related Subject Headings

  • Vascular Endothelial Growth Factor A
  • Real-Time Polymerase Chain Reaction
  • RNA, Messenger
  • Humans
  • Female
  • Enzyme-Linked Immunosorbent Assay
  • Endothelium, Vascular
  • Coculture Techniques
  • Cell Proliferation
  • Cell Line, Tumor
 

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Buchanan, C. F., Szot, C. S., Wilson, T. D., Akman, S., Metheny-Barlow, L. J., Robertson, J. L., … Rylander, M. N. (2012). Cross-talk between endothelial and breast cancer cells regulates reciprocal expression of angiogenic factors in vitro. J Cell Biochem, 113(4), 1142–1151. https://doi.org/10.1002/jcb.23447
Buchanan, Cara F., Christopher S. Szot, Tia D. Wilson, Steven Akman, Linda J. Metheny-Barlow, John L. Robertson, Joseph W. Freeman, and Marissa Nichole Rylander. “Cross-talk between endothelial and breast cancer cells regulates reciprocal expression of angiogenic factors in vitro.J Cell Biochem 113, no. 4 (April 2012): 1142–51. https://doi.org/10.1002/jcb.23447.
Buchanan CF, Szot CS, Wilson TD, Akman S, Metheny-Barlow LJ, Robertson JL, et al. Cross-talk between endothelial and breast cancer cells regulates reciprocal expression of angiogenic factors in vitro. J Cell Biochem. 2012 Apr;113(4):1142–51.
Buchanan, Cara F., et al. “Cross-talk between endothelial and breast cancer cells regulates reciprocal expression of angiogenic factors in vitro.J Cell Biochem, vol. 113, no. 4, Apr. 2012, pp. 1142–51. Pubmed, doi:10.1002/jcb.23447.
Buchanan CF, Szot CS, Wilson TD, Akman S, Metheny-Barlow LJ, Robertson JL, Freeman JW, Rylander MN. Cross-talk between endothelial and breast cancer cells regulates reciprocal expression of angiogenic factors in vitro. J Cell Biochem. 2012 Apr;113(4):1142–1151.
Journal cover image

Published In

J Cell Biochem

DOI

EISSN

1097-4644

Publication Date

April 2012

Volume

113

Issue

4

Start / End Page

1142 / 1151

Location

United States

Related Subject Headings

  • Vascular Endothelial Growth Factor A
  • Real-Time Polymerase Chain Reaction
  • RNA, Messenger
  • Humans
  • Female
  • Enzyme-Linked Immunosorbent Assay
  • Endothelium, Vascular
  • Coculture Techniques
  • Cell Proliferation
  • Cell Line, Tumor