Comparative proteomics of soluble factors secreted by human breast adipose tissue from tumor and normal breasts

VIRGINIA PISTONE CREYDT; LUCIANO VELLÓN; RUBÉN DRESZMAN; MARÍA LUJÁN CROSBIE; MARÍA PAULA MARKS; JUAN C. CALVO; JUDITH TONEATTO; ALBERTO GUTIÉRREZ; RUBÉN CARÓN; NATALIA SANTISO; EDUARDO CALLEGARI; MARÍA BELÉN HAPÓN; ALICIA RITA AMATO; ANABELA URSINO; SABRINA JOHANNA FLETCHER

Congreso; LXII Reunión Científica Anual de la Sociedad Argentina de Investigación Clínica (SAIC); 2017

Epithelial-stromal cell interaction is a crucial factor in cancer progression. In the breast, adipose tissue is the main stromal component. We have previously demonstrated that conditioned media (CMs) from human adipose tissue explants of tumor breasts (hATT) differentially regulate proliferation, adhesion and migration of breast cancer epithelial cells compared with CMs from human adipose tissue explants of normal breasts (hATN). Now, we intend to identify the proteins present in the CMs. For this, we separated in polyacrylamide gels proteins in the CMs ?hATT (n=6) and ?hATN (n=3). Aliquots from these CMs were analyzed by means of 2D nano-LC-MS/MS (mass spectrometry). The data was analyzed using ProteoIQ (Premier Biosoft) and FunRich softwares. In addition, CMs ?hATT (n=6) and -hATN (n=5) were assayed using a 42 Cytokine Antibody Array. We found that CMs-hATT present more protein diversity than CMs-hATN. Moreover, CMs-hATT expressed greater amount of proteins involved in biological processes such as signal transduction and cell communication; energy metabolism; cell growth; and immune response. Specifically, levels of apolipoprotein AI and AII (lipid metabolic processes), C3 complement factor (immune system) and vimentin and desmin (mesenchymal cells glycoprotein related to an invasive breast cancer phenotype) were significantly increased in CMs-hATT vs. CMs-hATN (cut-off 5-fold change). Also, a multivariate discriminant analysis of the cytokines detected by the array showed that IL-6, MCP-2 and GRO cytokines are sufficient and necessary to differentiate CMs-hATT from CMs-hATN. In addition, this analysis showed that the levels of these three cytokines taken together correlate with tumor stage of CMs-hATT and with BMI of CMs-hATN. These results allowed us to identify proteins potentially responsible for the observed effects, and let us proposed stromal IL-6, MCP-2 and GRO as potential markers of the stage of the disease.