title: 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.
abstract: 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.
authors.affiliation: Department of Gynecology of The Federal University of São Paulo, Brazil.|Charitable Association of Blood Collection - COLSAN, São Paulo, SP, Brazil.|Cell and Molecular Biology Laboratory, Center for Nuclear Energy in Agriculture CENA, University of São Paulo USP, Piracicaba, SP, Brazil.
authors.forename: Sheila Siqueira
authors.initials: SS
authors.lastname: Andrade
authors.affiliation: Department of Gynecology of The Federal University of São Paulo, Brazil.
authors.forename: Joana Tomomi
authors.initials: JT
authors.lastname: Sumikawa
authors.affiliation: Department of Biochemistry of The Federal University of São Paulo, Brazil.
authors.forename: Eloísa Dognani
authors.initials: ED
authors.lastname: Castro
authors.affiliation: Department of Biochemistry of The Federal University of São Paulo, Brazil.
authors.forename: Fabricio Pereira
authors.initials: FP
authors.lastname: Batista
authors.affiliation: Department of Biochemistry of The Federal University of São Paulo, Brazil.
authors.forename: Edgar
authors.initials: E
authors.lastname: Paredes-Gamero
authors.affiliation: Department of Biophysics of The Federal University of São Paulo, Brazil.
authors.forename: Lilian Carolina
authors.initials: LC
authors.lastname: Oliveira
authors.affiliation: Department of Gynecology of The Federal University of São Paulo, Brazil.
authors.forename: Izabel Monastério
authors.initials: IM
authors.lastname: Guerra
authors.affiliation: Department of Biochemistry of The Federal University of São Paulo, Brazil.
authors.forename: Giovani Bravin
authors.initials: GB
authors.lastname: Peres
authors.affiliation: Department of Biochemistry of The Federal University of São Paulo, Brazil.
authors.forename: Renan Pelluzzi
authors.initials: RP
authors.lastname: Cavalheiro
authors.affiliation: Department of Biophysics of The Federal University of São Paulo, Brazil.
authors.forename: Luiz
authors.initials: L
authors.lastname: Juliano
authors.affiliation: Department of Gynecology of The Federal University of São Paulo, Brazil.
authors.forename: Afonso Pinto
authors.initials: AP
authors.lastname: Nazário
authors.affiliation: Department of Gynecology of The Federal University of São Paulo, Brazil.
authors.forename: Gil
authors.initials: G
authors.lastname: Facina
authors.affiliation: Cell and Molecular Biology Laboratory, Center for Nuclear Energy in Agriculture CENA, University of São Paulo USP, Piracicaba, SP, Brazil.
authors.forename: Siu Mui
authors.initials: SM
authors.lastname: Tsai
authors.affiliation: Department of Biochemistry of The Federal University of São Paulo, Brazil.
authors.forename: Maria Luiza Vilela
authors.initials: ML
authors.lastname: Oliva
authors.affiliation: Department of Gynecology of The Federal University of São Paulo, Brazil.|Charitable Association of Blood Collection - COLSAN, São Paulo, SP, Brazil.
authors.forename: Manoel João Batista Castello
authors.initials: MJ
authors.lastname: Girão
mesh_terms: D001792:Blood Platelets / Q000473:pathology*; D001943:Breast Neoplasms / Q000098:blood supply / Q000473:pathology; D018450:Disease Progression; D004847:Epithelial Cells / Q000473:pathology; D005260:Female; D005337:Fibrin / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D009389:Neovascularization, Pathologic / Q000473:pathology; D053657:Platelet-Rich Plasma / Q000166:cytology*; D017154:Stromal Cells / Q000473:pathology; D059016:Tumor Microenvironment
chemical_list: D005337:Fibrin
keywords: breast cancer; cancer; platelet-rich plasma; platelets; tumor microenvironment