PMID- 26577851 OWN - NLM STAT- MEDLINE DCOM- 20170201 LR - 20220330 IS - 1423-0380 (Electronic) IS - 1010-4283 (Linking) VI - 37 IP - 4 DP - 2016 Apr TI - TNF-alpha increases the membrane expression of the chemokine receptor CCR6 in thyroid tumor cells, but not in normal thyrocytes: potential role in the metastatic spread of thyroid cancer. PG - 5569-75 LID - 10.1007/s13277-015-4418-7 [doi] AB - The chemokine receptor CCR6, selectively bound by CCL20, is involved in the metastatic spread of cancer cells. Tumor necrosis factor-alpha (TNF-alpha) displays a complex pro-tumorigenic actions, but it is unknown whether this cytokine could modulate the expression of chemokine receptors in thyroid tumors. The membrane expression of CCR6 was assessed by flow cytometry and immunofluorescence, in primary cultures of normal human thyroid (NHT) cells and in thyroid cancer cell lines (TPC-1 and BCPAP), both in basal conditions and after stimulation with TNF-alpha. In basal conditions, CCR6+ cells were virtually absent in NHT cells (0.4 +/- 0.4 %), while they were detected in TPC-1 (23.6 +/- 6.6 %) and in BCPAP (12.9 +/- 9.4 %) tumor cells (ANOVA F: 10.534; p < 0.005). The incubation with TNF-alpha significantly increased the percentage of CCR6+ cells in TPC-1 (23.6 +/- 6.6 % vs. 33.1 +/- 8.7; p < 0.033) and in BCPAP (12.9 +/- 9.4 % vs. 18.1 +/- 11.5; p < 0.030), but not in NHT (0.4 +/- 0.4 % vs. 0.2 +/- 0.3; NS) cells. The magnitude of the TNF-alpha effect was similar for TPC-1 and BCPAP ( approximately 40 % vs. baseline) cells. TPC-1 cells were characterized by a greater amount of CCR6 per cell as compared with BCPAP cells, both in basal conditions (148.3 +/- 33.7 fluorescence intensity vs. 102.5 +/- 22.1 p < 0.016) and after TNF-alpha stimulation (147.8 +/- 46.3 fluorescence intensity vs. 95.3 +/- 18.5; p < 0.025). Cell migration assays showed that TNF-alpha treatment significantly increased the rate of migrated cells in those cells in which it also increased the membrane expression of CCR6 (TPC-1 and BCPAP) as compared to basal condition (p < 0.05 for both TPC-1 and BCPAP cells). No effect was observed in NHT cells in which TNF-alpha stimulation had no effect in terms of CCR6 expression. We first report that TNF-alpha enhances the expression of CCR6 in thyroid tumor cells, thus providing evidence that TNF-alpha increases the metastatic potential of thyroid tumors. FAU - Coperchini, Francesca AU - Coperchini F AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Pignatti, Patrizia AU - Pignatti P AD - Allergy and Immunology Unit, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Carbone, Andrea AU - Carbone A AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Bongianino, Rossana AU - Bongianino R AD - Molecular Cardiology Unit, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Di Buduo, Christian A AU - Di Buduo CA AD - Department of Molecular Medicine, University of Pavia, Pavia, Italy. AD - Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Policlinico San Matteo Foundation, Pavia, Italy. FAU - Leporati, Paola AU - Leporati P AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Croce, Laura AU - Croce L AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Magri, Flavia AU - Magri F AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. FAU - Balduini, Alessandra AU - Balduini A AD - Department of Molecular Medicine, University of Pavia, Pavia, Italy. AD - Biotechnology Research Laboratories, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Policlinico San Matteo Foundation, Pavia, Italy. AD - Department of Biomedical Engineering, Tufts University, Medford, MA, USA. FAU - Chiovato, Luca AU - Chiovato L AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. luca.chiovato@fsm.it. FAU - Rotondi, Mario AU - Rotondi M AD - Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, Fondazione Salvatore Maugeri I.R.C.C.S., Via Maugeri 10, 27100, Pavia, Italy. LA - eng PT - Journal Article DEP - 20151117 PL - Netherlands TA - Tumour Biol JT - Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine JID - 8409922 RN - 0 (CCL20 protein, human) RN - 0 (CCR6 protein, human) RN - 0 (Chemokine CCL20) RN - 0 (Receptors, CCR6) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Cell Line, Tumor MH - Cell Movement/genetics MH - Chemokine CCL20/genetics/metabolism MH - Gene Expression Regulation, Neoplastic/drug effects MH - Humans MH - Neoplasm Invasiveness/*genetics MH - Neoplasm Metastasis MH - Receptors, CCR6/*biosynthesis/genetics MH - Thyroid Epithelial Cells/metabolism/pathology MH - Thyroid Gland/cytology/drug effects/metabolism MH - Thyroid Neoplasms/*genetics/pathology MH - Tumor Necrosis Factor-alpha/administration & dosage/*genetics OTO - NOTNLM OT - CCR6 OT - Chemokine receptor OT - Thyroid cancer OT - Thyroid cancer cell lines OT - Tumor necrosis factor-alpha EDAT- 2015/11/19 06:00 MHDA- 2017/02/02 06:00 CRDT- 2015/11/19 06:00 PHST- 2015/08/25 00:00 [received] PHST- 2015/11/10 00:00 [accepted] PHST- 2015/11/19 06:00 [entrez] PHST- 2015/11/19 06:00 [pubmed] PHST- 2017/02/02 06:00 [medline] AID - 10.1007/s13277-015-4418-7 [pii] AID - 10.1007/s13277-015-4418-7 [doi] PST - ppublish SO - Tumour Biol. 2016 Apr;37(4):5569-75. doi: 10.1007/s13277-015-4418-7. Epub 2015 Nov 17.