PDE5 expression in human thyroid tumors and effects of PDE5 inhibitors on growth and migration of cancer cells
- 479 Downloads
Recent studies have revealed in normal thyroid tissue the presence of the transcript of several phosphodiesterases (PDEs), enzymes responsible for the hydrolysis of cyclic nucleotides. In this work, we analyzed the expression of PDE5 in a series of human papillary thyroid carcinomas (PTCs) presenting or not BRAF V600E mutation and classified according to ATA risk criteria. Furthermore, we tested the effects of two PDE5 inhibitors (sildenafil, tadalafil) against human thyroid cancer cells. PDE5 gene and protein expression were analyzed in two different cohorts of PTCs by real-time PCR using a TaqMan micro-fluid card system and Western blot. MTT and migration assay were used to evaluate the effects of PDE5 inhibitors on proliferation and migration of TPC-1, BCPAP, and 8505C cells. In a first series of 36 PTCs, we found higher expression levels of PDE5A in tumors versus non-tumor (normal) tissues. PTCs with BRAF mutation showed higher levels of mRNA compared with those without mutation. No significant differences were detected between subgroups with low and intermediate ATA risk. Upregulation of PDE5 was also detected in tumor tissue proteins. Similar results were obtained analyzing the second cohort of 50 PTCs. Moreover, all tumor tissues with high PDE5 levels showed reduction of Thyroglobulin, TSH receptor, Thyroperoxidase, and NIS transcripts. In thyroid cancer cells in vitro, sildenafil and tadalafil determined a reduction of proliferation and cellular migration. Our findings demonstrate for the first time an overexpression of PDE5 in PTCs, and the ability of PDE5 inhibitors to block the proliferation of thyroid cancer cells in culture, therefore, suggesting that specific inhibition of PDE5 may be proposed for the treatment of these tumors.
KeywordsPapillary thyroid carcinoma Phosphodiesterases BRAF Thyroid cancer cells
This work was supported by Fondazione Umberto Di Mario.
This work is funded by grant to MC (MIUR: Grant RBFR12FI27_003).
Conflict of interest
The authors declare that they have no conflict of interest.
- 2.C.K. Jung, M.P. Little, J.H. Lubin, A.V. Brenner, S.A. Wells Jr, A.J. Sigurdson, Y.E. Nikiforov, The increase in thyroid cancer incidence during the last four decades is accompanied by a high frequency of BRAF mutations and a sharp increase in RAS mutations. J. Clin. Endocrinol. Metab. 99(2), E276–E285 (2014)PubMedCentralCrossRefPubMedGoogle Scholar
- 14.L.G. Bazzara, M.L. Vélez, M.E. Costamagna, A.M. Cabanillas, L. Fozzatti, A.M. Lucero, C.G. Pellizas, A.M. Masini-Repiso, Nitric oxide/cGMP signaling inhibits TSH-stimulated iodide uptake and expression of thyroid peroxidase and thyroglobulin mRNA in FRTL-5 thyroid cells. Thyroid 17(8), 717–727 (2007)CrossRefPubMedGoogle Scholar
- 18.D.S. Cooper, G.M. Doherty, B.R. Haugen, R.T. Kloos, S.L. Lee, S.J. Mandel, E.L. Mazzaferri, B. McIver, F. Pacini, M. Schlumberger, S.I. Sherman, D.L. Steward, R.M. Tuttle, Revised American Thyroid Association management guidelines for patients with thyroid nodules and differentiated thyroid cancer. Thyroid 19, 1167–1214 (2009)CrossRefPubMedGoogle Scholar
- 19.C. Durante, G. Tallini, E. Puxeddu, M. Sponziello, S. Moretti, C. Ligorio, A. Cavaliere, K.J. Rhoden, A. Verrienti, M. Maranghi, L. Giacomelli, D. Russo, S. Filetti, BRAF(V600E) mutation and expression of proangiogenic molecular markers in papillary thyroid carcinomas. Eur. J. Endocrinol. 165(3), 455–463 (2011)CrossRefPubMedGoogle Scholar
- 20.M.L. Sponziello, E. Lavarone, E. Pegolo, C. Di Loreto, C. Puppin, M.A. Russo, R. Bruno, S. Filetti, C. Durante, D. Russo, A. Di Cristofano, G. Damante, Molecular differences between human thyroid follicular adenoma and carcinoma revealed by analysis of a murine model of thyroid cancer. Endocrinology 154, 3043–3053 (2013)PubMedCentralCrossRefPubMedGoogle Scholar
- 21.M.L. Sponziello, R. Bruno, C. Durante, M. D’Agostino, R. Corradino, P. Giannasio, E. Ciociola, E. Ferretti, M. Maranghi, A. Verrienti, G. De Toma, S. Filetti, D. Russo, Growth factor receptors gene expression and Akt phosphorylation in benign human thyroid nodules are unaffected by chronic thyrotropin suppression. Horm. Metab. Res. 43(1), 22–25 (2011)CrossRefPubMedGoogle Scholar
- 22.R.E. Schweppe, J.P. Klopper, C. Korch, U. Puqazhenthi, M. Benezra, J.A. Knauf, J.A. Fagin, L.A. Marlow, J.A. Copland, R.C. Smallridge, B.R. Haugen, Deoxyribonucleic acid profiling analysis of 40 human thyroid cancer cell lines reveals cross-contamination resulting in cell line redundancy and misidentification. J. Clin. Endocrinol. Metab. 93(11), 4331–4341 (2008)PubMedCentralCrossRefPubMedGoogle Scholar
- 24.M. D’Agostino, P. Voce, M. Celano, M. Sponziello, S. Moretti, V. Maggisano, A. Verrienti, C. Durante, S. Filetti, E. Puxeddu, D. Russo, Sunitinib exerts only limited effects on the proliferation and differentiation of anaplastic thyroid cancer cells. Thyroid 22, 138–144 (2012)CrossRefPubMedGoogle Scholar
- 25.V. Maggisano, C. Puppin, M. Celano, M. D’Agostino, M. Sponziello, S. Micali, M. Navarra, G. Damante, S. Filetti, D. Russo, Cooperation of histone deacetylase inhibitors SAHA and valproic acid in promoting sodium/iodide symporter expression and function in rat Leydig testicular carcinoma cells. Endocrine 45(1), 148–152 (2014)CrossRefPubMedGoogle Scholar
- 30.A.L. Galrão, A.K. Sodré, R.Y. Camargo, C.U. Friguglietti, M.A. Kulcsar, E.U. Lima, G. Medeiros-Neto, I.G. Rubio, Methylation levels of sodium-iodide symporter (NIS) promoter in benign and malignant thyroid tumors with reduced NIS expression. Endocrine 43(1), 225–229 (2013)CrossRefPubMedGoogle Scholar
- 31.C. Puppin, F. D’Aurizio, A.V. D’Elia, L. Cesaratto, G. Tell, D. Russo, S. Filetti, E. Ferretti, E. Tosi, T. Mattei, A. Pianta, L. Pellizzari, G. Damante, Effects of histone acetylation on sodium iodide symporter promoter and expression of thyroid-specific transcription factors. Endocrinology 146(9), 3967–3974 (2005)CrossRefPubMedGoogle Scholar