Comparative proteomics of soluble factors secreted by human breast adipose tissue from tumor and normal breast
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Sabrina Johanna Fletcher1, María Belén Hapon9, Eduardo A. Callegari3, María Luján Crosbie4, Natalia Santiso4, Anabela Ursino4, Alicia Rita Amato4, Alberto Gutiérrez4, Paula Alejandra Sacca1, Rubén Dreszman5, Adriana Pérez6, Rubén Walter Carón2, Juan Carlos Calvo1,7 and Virginia Pistone-Creydt1,2,8
1Laboratorio de Química de Proteoglicanos y Matriz Extracelular, Instituto de Biología y Medicina Experimental (IBYME), Buenos Aires, Argentina
2Laboratorio de Hormonas y Biología del Cáncer, Instituto de Medicina y Biología Experimental de Cuyo (IMBECU), Centro Científico y Tecnológico Mendoza, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Mendoza, Argentina
3University of South Dakota, Sanford School of Medicine, Vermillion, South Dakota, USA
4Sección de Patología Mamaria, Servicio de Ginecología, Complejo Médico Policial “Churruca-Visca”, Buenos Aires, Argentina
5Clínica de Microcirugía, Buenos Aires, Argentina
6Departamento de Ecología, Genética y Evolución, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
7Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
8Departamento de Fisiología, Universidad Nacional de Cuyo, Facultad de Ciencias Médicas, Mendoza, Argentina
9Laboratorio de Reproducción y Lactancia, Instituto de Medicina y Biología Experimental de Cuyo (IMBECU), Centro Científico y Tecnológico Mendoza, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Mendoza, Argentina
Virginia Pistone-Creydt, email: firstname.lastname@example.org
Keywords: human breast cancer; adipose tissue; epithelial-stromal interaction; proteomics analysis; tumor microenvironment
Received: December 30, 2017 Accepted: June 24, 2018 Published: July 24, 2018
Tumor progression depends on the tumor-stroma interaction. In the breast, adipose tissue is the predominant stromal type. We have previously demonstrated that conditioned media (CMs) from explants of human adipose tissue of tumor breasts (hATT) increase proliferation and migration of breast cancer epithelial cells when compared to human adipose tissue from normal breasts (hATN). In this work, we aim to identify specific proteins and molecular/biological pathways associated with the secretion profile of hATT and hATN explants.
hATT-CMs and hATN-CMs were separated by SDS-PAGE and analyzed by means of two-dimensional nano-liquid chromatography-mass spectrometry. The data was analyzed using ProteoIQ and FunRich software. In addition, 42 cytokines from hATT-CMs and hATN-CMs were assayed by a protein antibody assay. Compared to hATN-CMs, hATT-CMs showed greater protein diversity. We found that hATT-CMs presented a greater amount of proteins related to complement system activity, metabolism and immune system, as well as proteins involved in a variety of biological processes such as signal transduction and cell communication. Specifically, apolipoprotein AI and AII, complement component 3, and vimentin and desmin were significantly increased in hATT-CMs versus hATN-CMs. Moreover, a multivariate discriminant analysis of the cytokines detected by the array showed that IL-6, MCP-2 and GRO cytokines were sufficient and necessary to differentiate hATT-CMs from hATN-CMs. This analysis also showed that the levels of these three cytokines, taken together, correlated with stage and histological grade of the tumor in the hATT-CMs group, and with body mass index in the hATN-CMs group.
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