title: Multicellular spheroids from normal and neoplastic thyroid tissues as a suitable model to test the effects of multikinase inhibitors.
abstract: Multicellular three-dimensional (3D) spheroids represent an experimental model that is intermediate in its complexity between monolayer cultures and patients' tumor. In the present study, we characterize multicellular spheroids from papillary (PTC) and follicular (FTC) thyroid cancers and from the corresponding normal tissues. We show that these 3D structures well recapitulate the features of the original tissues, in either the differentiated and "stem-like" components. As a second step, we were aimed to test the effects of a small multikinase inhibitor, SP600125 (SP), previously shown to efficiently induce cell death in undifferentiated thyroid cancer monolayer cultures. We demonstrate the potent effect of SP on cell growth and survival in our 3D multicellular cultures. SP exerts its main effects through direct and highly significant inhibition of the ROCK pathway, known to be involved in the regulation of cell migration and β-catenin turnover. Consistently, SP treatment resulted in a significant decrease in β-catenin levels with respect to basal conditions in tumor but not in normal spheroids, indicating that the effect is promisingly selective on tumor cells.In conclusion, we provide the morphological and molecular characterization of thyroid normal and tumor spheroids. In this 3D model we tested in vitro the effects of the multikinase inhibitor SP and further characterized its mechanism of action in both normal and tumor spheroids, thus making it an ideal candidate for developing new drugs against thyroid cancer.
authors.affiliation: Endocrine Unit, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.|Department of Pathophysiology and Transplantation, University of Milan, 20122 Milan, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Cirello
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Vaira
authors.affiliation: Laboratory of Endocrine and Metabolic Research, Istituto Auxologico Italiano IRCCS, 20149 Milan, Italy.
authors.forename: Elisa Stellaria
authors.initials: ES
authors.lastname: Grassi
authors.affiliation: Laboratory of Endocrine and Metabolic Research, Istituto Auxologico Italiano IRCCS, 20149 Milan, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Vezzoli
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.
authors.forename: Dario
authors.initials: D
authors.lastname: Ricca
authors.affiliation: Endocrine Unit, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.|Department of Clinical Sciences and Community Health, University of Milan, 20122 Milan, Italy.
authors.forename: Carla
authors.initials: C
authors.lastname: Colombo
authors.affiliation: Department of Pathophysiology and Transplantation, University of Milan, 20122 Milan, Italy.|Division of Pathology, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.
authors.forename: Silvano
authors.initials: S
authors.lastname: Bosari
authors.affiliation: Endocrine Surgery Unit, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Vicentini
authors.affiliation: Laboratory of Endocrine and Metabolic Research, Istituto Auxologico Italiano IRCCS, 20149 Milan, Italy.|Department of Clinical Sciences and Community Health, University of Milan, 20122 Milan, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Persani
authors.affiliation: Division of Pathology, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.|Department of Biomedical, Surgical and Dental Sciences, University of Milan, 20122 Milan, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferrero
authors.affiliation: Endocrine Unit, Fondazione IRCCS Ca' Granda, 20122 Milan, Italy.|Department of Pathophysiology and Transplantation, University of Milan, 20122 Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Fugazzola
mesh_terms: D018263:Adenocarcinoma, Follicular / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000873:Anthracenes / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002291:Carcinoma, Papillary / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D016022:Case-Control Studies; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D058990:Molecular Targeted Therapy; D010641:Phenotype; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011494:Protein Kinases / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D018874:Spheroids, Cellular; D000077273:Thyroid Cancer, Papillary; D013961:Thyroid Gland / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D013964:Thyroid Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology; D014407:Tumor Cells, Cultured; D051176:beta Catenin / Q000378:metabolism; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D000873:Anthracenes; D000970:Antineoplastic Agents; C495270:CTNNB1 protein, human; D047428:Protein Kinase Inhibitors; D051176:beta Catenin; C432165:pyrazolanthrone; D011494:Protein Kinases; D054460:rho-Associated Kinases
keywords: ROCK; SP600125; multi-tyrosine kinase inhibitors; multicellular spheroids; thyroid cancer