Interface between breast cancer cells and the tumor microenvironment using platelet-rich plasma to promote tumor angiogenesis - influence of platelets and fibrin bundles on the behavior of breast tumor cells
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Sheila Siqueira Andrade1,4,5, Joana Tomomi Sumikawa1, Eloísa Dognani Castro2, Fabricio Pereira Batista2, Edgar Paredes-Gamero2, Lilian Carolina Oliveira3, Izabel Monastério Guerra1, Giovani Bravin Peres2, Renan Pelluzzi Cavalheiro2, Luiz Juliano3, Afonso Pinto Nazário1, Gil Facina1, Siu Mui Tsai5, Maria Luiza Vilela Oliva2, Manoel João Batista Castello Girão1,4
1Department of Gynecology of The Federal University of São Paulo, Brazil
2Department of Biochemistry of The Federal University of São Paulo, Brazil
3Department of Biophysics of The Federal University of São Paulo, Brazil
4Charitable Association of Blood Collection – COLSAN, São Paulo, SP, Brazil
5Cell and Molecular Biology Laboratory, Center for Nuclear Energy in Agriculture CENA, University of São Paulo USP, Piracicaba, SP, Brazil
Sheila Siqueira Andrade, email: [email protected]
Keywords: platelets, breast cancer, platelet-rich plasma, cancer, tumor microenvironment
Received: December 14, 2016 Accepted: January 24, 2017 Published: February 07, 2017
Cancer progression is associated with an evolving tissue interface of direct epithelial-tumor microenvironment interactions. In biopsies of human breast tumors, extensive alterations in molecular pathways are correlated with cancer staging on both sides of the tumor-stroma interface. These interactions provide a pivotal paracrine signaling to induce malignant phenotype transition, the epithelial-mesenchymal transition (EMT). We explored how the direct contact between platelets-fibrin bundles primes metastasis using platelet-rich plasma (PRP) as a source of growth factors and mimics the provisional fibrin matrix between actively growing breast cancer cells and the tumor stroma. We have demonstrated PRP functions, modulating cell proliferation that is tumor-subtype and cancer cell-type-specific. Epithelial and stromal primary cells were prepared from breast cancer biopsies from 21 women with different cancer subtypes. Cells supplemented with PRP were immunoblotted with anti-phospho and total Src-Tyr-416, FAK-Try-925, E-cadherin, N-cadherin, TGF-β, Smad2, and Snail monoclonal antibodies. Breast tumor cells from luminal B and HER2 subtypes showed the most malignant profiles and the expression of thrombin and other classes of proteases at levels that were detectable through FRET peptide libraries. The angiogenesis process was investigated in the interface obtained between platelet-fibrin-breast tumor cells co-cultured with HUVEC cells. Luminal B and HER2 cells showed robust endothelial cell capillary-like tubes ex vivo. The studied interface contributes to the attachment of endothelial cells, provides a source of growth factors, and is a solid substrate. Thus, replacement of FBS supplementation with PRP supplementation represents an efficient and simple approach for mimicking the real multifactorial tumor microenvironment.
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