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

FLETCHER, S.; HAPON, M.B.; MARKS, M.P.; URSINO, A.; SANTISO, N.; CROSBIE, M.L.; AMATO, A.R.; GUTIÉRREZ, A.; DRESZMAN, R.; CALLEGARI, E; TONEATTO, J.; VELLÓN, L.; CARON, R.W.; CALVO, J.C.; PISTONE-CREYDT, V.

Buenos Aires

Congreso; Reunión conjunta de sociedades de Biociencias; 2017

Sociedad Argentina de Investigación Clínica

Epithelial-stromal cell interaction is a crucial factor in cancer progressionAdipose tissue is the main stromal component of breasts.We have previously demonstrated that conditioned media (CMs)from human adipose tissue explants of tumor breasts (hATT) differentiallyregulate proliferation, adhesion and migration of breast cancerepithelial cells compared with CMs from human adipose tissueexplants of normal breasts (hATN). Now, we intend to identify theproteins present in those CMs. For this, we separated in polyacrylamidegels proteins in CMs ?hATT (n=6) and ?hATN (n=3). Aliquotsfrom these CMs were analyzed by means of 2D nano-LC-MS/MS(mass spectrometry). The data was analyzed using ProteoIQ (PremierBiosoft) 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 CMshATN.Moreover, CMs-hATT expressed greater amount of proteinsinvolved in biological processes such as signal transduction and cellcommunication; energy metabolism; cell growth; and immune response.Specifically, levels of apolipoprotein AI and AII (lipid metabolicprocesses), C3 complement factor (immune system) and vimentinand desmin (mesenchymal cells glycoprotein related to an invasivebreast cancer phenotype) were significantly increased in CMs-hATTvs. CMs-hATN (cut-off 5-fold change). Furthermore, a multivariatediscriminant analysis of the cytokines detected by the array showedthat IL-6, MCP-2 and GRO cytokines are sufficient and necessaryto differentiate CMs-hATT from CMs-hATN. In addition, this analysisshowed that the levels of these three cytokines taken together correlatewith tumor stage of CMs-hATT and with BMI of CMs-hATN.These results allowed us to identify proteins potentially responsiblefor the observed effects, and let us proposed stromal IL-6, MCP-2and GRO as potential markers of the stage of the disease.Keywords: breast cancer, adipose tissue, epithelial-stromal interaction,proteomics analysis.