Thyroid tumor cells-fibroblasts interaction produces extracelular vesicles involved in thyroid tumor aggressiveness

BRAVO-MIANA R; NEGRETTI-BORGA D; SOLER MF; DE PAUL AL; GILARDONI MB; MONTESINOS, MM; PELLIZAS CG; DONADIO, AC

Buenos Aires

Congreso; XVI Congreso Latinoamericano de Tiroides (LATS); 2019

Sociedad Latinoamericana de Tiroides (LATS)

Introduction: Carcinomas are complex societies of mutually interacting cells. Extracellular vesicles (EVs) mediate intercellularcommunication in the tumor microenvironment (TM). Metalloproteinases (MMPs) activity and Galectin-1 (Gal-1) expression in TMregulate tumor-stroma crosstalk leading to tumor progression and aggressiveness. We demonstrated that thyroid tumor cells- fibroblasts(Fb) interactions induce the secretion of MMPs to culture supernatants (CMs) and a migratory phenotype in tumor cells. Objectives:To characterise EVs released by Fb and thyroid cells, and their role mediating a thyroid tumor-promoting environment. Methods:Thyroid tumor cells (TPC-1, 8505c) or thyroid non-tumor cells (NThyOri) were co-cultured with normal human Fb as a simulation ofthe thyroid TM. EVs, obtained by ultracentrifugation of thyroid cells, Fb and thyroid cells-Fb CMs were characterised by TransmissionElectron Microscopy and Western blot. MMPs activity was analysed by zymography in EVs, and in CMs from Fb and NThyOri cellsincubated with EVs. Gal-1 expression was evaluated by RT-qPCR in EVs-stimulated Fb. Results: Isolated and co-cultured cellssecreted EVs to CMs. EVs have the characteristic cup-shaped morphology, size (<150 nm), and CD81 and CD63 expression describedfor exosomes. EVs released by TPC-1-Fb co-cultures induced proMMP2 secretion and the significant MMP2 activation from Fb andNThyOri cells, as well as increased proMMP9 expression from NThyOri cells. Gal-1 mRNA expression increased in Fb stimulated withEVs from TPC-1-Fb co-cultures. Conclusion: These findings underpin the roles of: 1) the interaction of thyroid tumor cells and Fb inthe promotion of tumor aggressiveness, and 2) EVs as contributors in this crosstalk.