Andrade Sheila Siqueira, Sumikawa Joana Tomomi, Castro Eloísa Dognani, Batista Fabricio Pereira, Paredes-Gamero Edgar, Oliveira Lilian Carolina, Guerra Izabel Monastério, Peres Giovani Bravin, Cavalheiro Renan Pelluzzi, Juliano Luiz, Nazário Afonso Pinto, Facina Gil, Tsai Siu Mui, Oliva Maria Luiza Vilela, Girão Manoel João Batista Castello
Department of Gynecology of The Federal University of São Paulo, Brazil.
Charitable Association of Blood Collection - COLSAN, São Paulo, SP, Brazil.
Oncotarget. 2017 Mar 7;8(10):16851-16874. doi: 10.18632/oncotarget.15170.
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.
癌症进展与上皮细胞 - 肿瘤微环境直接相互作用不断演变的组织界面相关。在人类乳腺肿瘤活检中，肿瘤 - 基质界面两侧分子途径的广泛改变与癌症分期相关。这些相互作用提供了关键的旁分泌信号，以诱导恶性表型转变，即上皮 - 间质转化（EMT）。我们利用富含血小板的血浆（PRP）作为生长因子的来源，探讨了血小板 - 纤维蛋白束之间的直接接触如何引发转移，并模拟活跃生长的乳腺癌细胞与肿瘤基质之间的临时纤维蛋白基质。我们已经证明PRP具有功能，可调节肿瘤亚型和癌细胞类型特异性的细胞增殖。从21名患有不同癌症亚型的女性的乳腺癌活检中制备上皮和基质原代细胞。用抗磷酸化和总Src - Tyr - 416、FAK - Try - 925、E - 钙黏蛋白、N - 钙黏蛋白、TGF - β、Smad2和Snail单克隆抗体对补充有PRP的细胞进行免疫印迹分析。管腔B型和HER2亚型的乳腺肿瘤细胞显示出最恶性的特征，并且凝血酶和其他类别的蛋白酶的表达水平可通过FRET肽库检测到。在与HUVEC细胞共培养的血小板 - 纤维蛋白 - 乳腺肿瘤细胞之间获得的界面中研究血管生成过程。管腔B型和HER2细胞在体外显示出强大的内皮细胞毛细血管样管。所研究的界面有助于内皮细胞的附着，提供生长因子来源，并且是一个固体基质。因此，用PRP补充替代FBS补充是模拟真实多因素肿瘤微环境的一种有效且简单的方法。
