Bravo-Miana Rocío Del Carmen, Della Vedova Ana Belén, De Paul Ana Lucía, Remedi María Mónica, Guantay María Laura, Gilardoni Mónica Beatriz, Pellizas Claudia Gabriela, Donadio Ana Carolina
Universidad Nacional de Córdoba, Facultad de Ciencias Químicas, Departamento de Bioquímica Clínica, Córdoba, Argentina.
Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI-CONICET), Haya de la Torre y Medina Allende, Ciudad Universitaria, Córdoba (X5000HUA), Argentina.
Endocr Connect. 2020 Jun;9(6):506-518. doi: 10.1530/EC-20-0113.
Tumor-stroma crosstalk leads to a tumor-promoting microenvironment. In this milieu, extracellular vesicles (EVs) are protagonists in cell-cell communication. Despite thyroid cancer being the most common endocrine malignancy, the contribution of the tumor microenvironment to thyroid cancer progression is still largely underexplored. We focused on the role of thyroid tumor cell-fibroblast interaction and EVs as mediators of tumor-stroma interplay, in the promotion of thyroid tumor aggressiveness. Thyroid tumor (TPC-1, 8505c) or non-tumor thyroid cells (NThyOri) were co-cultured with human fibroblasts (Fb). Thyroid cell migration was investigated by the wound-healing assay and actin-network staining. Cell-CD147 expression was characterized by flow cytometry. EVs, obtained by ultracentrifugation of conditioned media (CMs), were characterized by transmission electron-microscopy and CD81 and CD147 expression. Metalloproteinases (MMPs) were evaluated by zymography in CMs. A migratory phenotype was triggered in thyroid tumor cells treated with CMs from Fb or from Fb-thyroid tumor cell co-cultures. Fb-thyroid cell co-cultures induced the secretion of proMMP9 and proMMP2 and led to a significant MMP2 activation in CMs. Fb, thyroid cells and Fb-thyroid cell co-cultures released EVs, and remarkably, EVs released by Fb-thyroid tumor cell co-cultures induced the secretion of proMMP2 and the expression of MMP2 from normal Fb. A significant CD147 expression was demonstrated in Fb-thyroid tumor cell-derived EVs. These findings reveal the role of Fb and thyroid tumor cell-Fb interaction in the promotion of a microenvironment suitable for thyroid tumor progression. Moreover, they highlight, for the first time, the role of thyroid tumor cell-Fb interaction in the production of specialized EVs.
肿瘤-基质相互作用导致促肿瘤微环境的形成。在这种环境中，细胞外囊泡（EVs）是细胞间通讯的主角。尽管甲状腺癌是最常见的内分泌恶性肿瘤，但肿瘤微环境对甲状腺癌进展的作用仍在很大程度上未被充分探索。我们聚焦于甲状腺肿瘤细胞与成纤维细胞的相互作用以及细胞外囊泡作为肿瘤-基质相互作用的介质在促进甲状腺肿瘤侵袭性方面的作用。将甲状腺肿瘤细胞（TPC-1、8505c）或非肿瘤甲状腺细胞（NThyOri）与人成纤维细胞（Fb）共培养。通过伤口愈合试验和肌动蛋白网络染色研究甲状腺细胞迁移。通过流式细胞术对细胞CD147表达进行表征。通过对条件培养基（CMs）进行超速离心获得细胞外囊泡，通过透射电子显微镜以及CD81和CD147表达对其进行表征。通过酶谱法在CMs中评估金属蛋白酶（MMPs）。用来自Fb或Fb-甲状腺肿瘤细胞共培养物的CMs处理的甲状腺肿瘤细胞引发了迁移表型。Fb-甲状腺细胞共培养诱导了前MMP9和前MMP2的分泌，并导致CMs中MMP2的显著激活。Fb、甲状腺细胞和Fb-甲状腺细胞共培养物释放细胞外囊泡，值得注意的是，Fb-甲状腺肿瘤细胞共培养物释放的细胞外囊泡诱导了前MMP2的分泌以及正常Fb中MMP2的表达。在Fb-甲状腺肿瘤细胞衍生的细胞外囊泡中证实了显著的CD147表达。这些发现揭示了Fb以及甲状腺肿瘤细胞与Fb的相互作用在促进适合甲状腺肿瘤进展的微环境中的作用。此外，它们首次突出了甲状腺肿瘤细胞与Fb的相互作用在产生特殊细胞外囊泡中的作用。
