Human thyroid cancer derived cell lines are widely used to study the mechanisms involved in thyroid carcinogenesis. However, there is limited availability of non cross-contaminated cancer cell lines derived from papillary thyroid carcinoma (PTC), and the B-CPAP cell line is one of the few such lines. B-CPAP cells have been genetically and cytogenetically well-characterized, but details of their stemness features remain uncertain. We broaden the functional and molecular profiles as well as the tumorigenic capacity of this cell line using functional assays (sphere-forming capacity and efficiency), assessing self-renewal and propagation efficiency, and testing in vivo tumorigenicity in Hsd:Athymic Nude-Foxn1nu mice. Expression of markers of stemness, differentiation, and epithelial mesenchymal transition were estimated at RNA and protein levels in adherent parental cells and sphere-forming cells. Protein expression of xenograft tumors was evaluated by immunohistochemistry. B-CPAP sphere-forming cells were able to form thyrospheres theoretically indefinitely in an appropriate serum-free medium, reverting to the adherent parental cell phenotype when cultured in differentiation medium. Different expression of ALDH1-A1 and CD44 stemness markers and TTF-1 and CK19 differentiation markers allowed discrimination between isolated sphere-forming cells and adherent parental cells, indicating that sphere-forming cells retained stem-like features. In keeping with these observations, tumorigenicity assays confirmed that, relative to parental adherent cells, thyrospheres had enhanced capacity to initiate xenograft tumors. Our findings may constitute a useful background to develop an in vitro model for assessing the origin and progression of papillary thyroid carcinoma bearing BRAFV600E and TERT promoter mutations

Thyrospheres from B-CPAP Cell Line with BRAF and TERT promoter mutations have Different Functional and Molecular Features than Parental Cells

GIOVANNONI, ROBERTO;
2017

Abstract

Human thyroid cancer derived cell lines are widely used to study the mechanisms involved in thyroid carcinogenesis. However, there is limited availability of non cross-contaminated cancer cell lines derived from papillary thyroid carcinoma (PTC), and the B-CPAP cell line is one of the few such lines. B-CPAP cells have been genetically and cytogenetically well-characterized, but details of their stemness features remain uncertain. We broaden the functional and molecular profiles as well as the tumorigenic capacity of this cell line using functional assays (sphere-forming capacity and efficiency), assessing self-renewal and propagation efficiency, and testing in vivo tumorigenicity in Hsd:Athymic Nude-Foxn1nu mice. Expression of markers of stemness, differentiation, and epithelial mesenchymal transition were estimated at RNA and protein levels in adherent parental cells and sphere-forming cells. Protein expression of xenograft tumors was evaluated by immunohistochemistry. B-CPAP sphere-forming cells were able to form thyrospheres theoretically indefinitely in an appropriate serum-free medium, reverting to the adherent parental cell phenotype when cultured in differentiation medium. Different expression of ALDH1-A1 and CD44 stemness markers and TTF-1 and CK19 differentiation markers allowed discrimination between isolated sphere-forming cells and adherent parental cells, indicating that sphere-forming cells retained stem-like features. In keeping with these observations, tumorigenicity assays confirmed that, relative to parental adherent cells, thyrospheres had enhanced capacity to initiate xenograft tumors. Our findings may constitute a useful background to develop an in vitro model for assessing the origin and progression of papillary thyroid carcinoma bearing BRAFV600E and TERT promoter mutations
Caria, P; Pillai, R; Dettori, T; Frau, D; Zavattari, P; Riva, G; Romano, G; Pani, F; Bentivegna, A; Giovannoni, Roberto; Pagni, F; Sogos, V; Vanni, R
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Descrizione: Caria P et al J Cancer 2017 EDITOR VERSION
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11568/945480
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