title: MAPK and SHH pathways modulate type 3 deiodinase expression in papillary thyroid carcinoma.
abstract: Type 3 deiodinase (DIO3, D3) is reactivated in human neoplasias. Increased D3 levels in papillary thyroid carcinoma (PTC) have been associated with tumor size and metastatic disease. The objective of this study is to investigate the signaling pathways involved in DIO3 upregulation in PTC. Experiments were performed in human PTC cell lines (K1 and TPC-1 cells) or tumor samples. DIO3 mRNA and activity were evaluated by real-time PCR and ion-exchange column chromatography respectively. Western blot analysis was used to determine the levels of D3 protein. DIO3 gene silencing was performed via siRNA transfection. DIO3 mRNA levels and activity were readily detected in K1 (BRAF(V6) (0) (0E)) and, at lower levels, in TPC-1 (RET/PTC1) cells (P<0.007 and P=0.02 respectively). Similarly, DIO3 mRNA levels were higher in PTC samples harboring the BRAF(V600E) mutation as compared with those with RET/PTC1 rearrangement or negative for these mutations (P<0.001). Specific inhibition of BRAF oncogene (PLX4032, 3 μM), MEK (U0126, 10-20 μM) or p38 (SB203580, 10-20 μM) signaling was associated with decreases in DIO3 expression in K1 and TPC-1 cells. Additionally, the blockage of the sonic hedgehog (SHH) pathway by cyclopamine (10  μM) resulted in markedly decreases in DIO3 mRNA levels. Interestingly, siRNA-mediated DIO3 silencing induced decreases on cyclin D1 expression and partial G1 phase cell cycle arrest, thereby downregulating cell proliferation. In conclusion, sustained activation of the MAPK and SHH pathways modulate the levels of DIO3 expression in PTC. Importantly, DIO3 silencing was associated with decreases in cell proliferation, thus suggesting a D3 role in tumor growth and aggressiveness.
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Mírian
authors.initials: M
authors.lastname: Romitti
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Simone Magagnin
authors.initials: SM
authors.lastname: Wajner
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Lucieli
authors.initials: L
authors.lastname: Ceolin
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Carla Vaz
authors.initials: CV
authors.lastname: Ferreira
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Rafaela Vanin Pinto
authors.initials: RV
authors.lastname: Ribeiro
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Helena Cecin
authors.initials: HC
authors.lastname: Rohenkohl
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Shana de Souto
authors.initials: Sde S
authors.lastname: Weber
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Patrícia Luciana da Costa
authors.initials: PL
authors.lastname: Lopez
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Cesar Seigi
authors.initials: CS
authors.lastname: Fuziwara
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil.
authors.forename: Edna Teruko
authors.initials: ET
authors.lastname: Kimura
authors.affiliation: Thyroid SectionEndocrine Division, Serviço de Endocrinologia, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2350, CEP 90035-003 Porto Alegre, RS, BrazilExperimental Research CenterHospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, BrazilDepartment of Cell and Developmental BiologyInstitute of Biomedical Sciences, Universidade de São Paulo, São Paulo, SP, Brazil almaia@ufrgs.br.
authors.forename: Ana Luiza
authors.initials: AL
authors.lastname: Maia
mesh_terms: D002277:Carcinoma / Q000235:genetics / Q000378:metabolism*; D002291:Carcinoma, Papillary; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics / Q000378:metabolism*; D048369:MAP Kinase Kinase 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009154:Mutation; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction; D000077273:Thyroid Cancer, Papillary; D013964:Thyroid Neoplasms / Q000235:genetics / Q000378:metabolism*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D053823:Hedgehog Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C509239:SHH protein, human; C117274:iodothyronine deiodinase type III; D007453:Iodide Peroxidase; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D048051:p38 Mitogen-Activated Protein Kinases; D048369:MAP Kinase Kinase 1; C482125:MAP2K1 protein, human
keywords: MAPK genetic alterations; Sonic Hedgehog pathway; type 3 deiodinase, papillary thyroid carcinoma