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Article in English | WPRIM | ID: wpr-928247


Objective This study was designed to determine the methylation profile of four CpGs and the genotypes of two CpG-SNPs located in promoter region of DIO2 in patients with Kashin-Beck disease (KBD). We also analyzed the interaction between the CpGs methylations and CpG-SNPs. Methods Whole blood specimens were collected from 16 KBD patients and 16 healthy subjects. Four CpGs and two CpG-SNPs in the promoter regions of DIO2 were detected using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The CpGs methylation levels were compared between samples from KBD patients and healthy subjects. The methylation levels were also analyzed in KBD patients with different CpG-SNP genotypes. Results The mRNA expression of DIO2 in whole blood of KBD patients was significnatly lower than in healthy controls (P <0.05). The methylation levels of DIO2-1_CpG_3 in KBD patients were significantly higher than those in healthy controls (P <0.05). The methylation levels of four CpGs were not significantly different between KBD patients and healthy controls. The methylation level of DIO2-1_CpG_3 in the promoter region of DIO2 in KBD patients with GA/AA genotype was significantly higher than that of KBD patients with GG genotype (P <0.05). Conclusion The methylation level of DIO2 increases in KBD patients. Similar trends exist in KBD carriers of variant genotypes of CpG-SNPs DIO2 rs955849187.

Humans , Case-Control Studies , Iodide Peroxidase/genetics , Kashin-Beck Disease/genetics , Methylation , Polymorphism, Single Nucleotide , Promoter Regions, Genetic
Medicina (B.Aires) ; 81(3): 346-358, jun. 2021. graf
Article in English | LILACS | ID: biblio-1346469


Abstract Hepatocellular carcinoma (HCC) is the most common primary liver tumor. Hexachlorobenzene (HCB) is an endocrine disruptor and a liver tumor promoter. Deregulation of thyroid hormone (TH) homeostasis may play a significant role in early neoplastic transformation. The aim of this study was to evaluate the relation between TH metabolism and the regulation of cell growth in an in vivo and in vitro model. We examined the role of transforming growth factor-β1 (TGF-β1) on TH deiodinase expression and hepatocyte proliferation. An initiation (DEN)/promotion (HCB) tumor model from rat liver and HepG2 cells were used. We evaluated PCNA, p21, p27, SMAD2/3, TGF-β1, deiodinase 1 (D1), D3, protein expression levels; D1 and D3 mRNA expression; TH and TGF-β1, D1, D3, and GST-P protein levels in focal/non-focal areas. In vivo, HCB decreased triiodothyronine (T3) and D1 mRNA levels and increased thyroxine (T4) and D3 mRNA levels in liver from DEN+HCB vs. DEN group. HCB increased protein levels from D3, TGF-β1, and PCNA and decreased D1 in focal-areas. In vitro, HCB increased PCNA, pSMAD 2/3, and TGF-β1 protein levels and mRNA expression and decreased p21 and p27 protein levels. Exogenous T3 treatment prevent HCB induced molecular alterations related to hepatocyte proliferation whereas T4 did not have any effect. These effects were prevented by using a TGF-β1 receptor II inhibitor. Results suggest that alteration of TH homeostasis, through D1 function, play a key role in hepatocyte proliferation and that TGF-β1-SMAD pathway is involved in this process confirming their role in early neoplastic transformation in HCC.

Resumen El hepatocarcinoma (HCC) es un tumor hepático primario. El hexaclorobenceno (HCB) es un disruptor endocrino y un promotor de tumores hepáticos. La desregulación de la homeostasis de las hormonas tiroideas (HT) puede ser un proceso importante para la transformación neoplásica temprana. Nuestro objetivo fue evaluar la relación entre el metabolismo de las HT y la regulación de la prolifera ción celular. Se utilizó un modelo tumoral de iniciación (DEN)/promoción (HCB) de hígado de rata (in vivo) (DEN/ HCB) y células HepG2 (in vitro). Evaluamos los niveles de PCNA, p21, p27, SMAD2/3, TGF-β1, D1, D3, ARNm de D1 y D3, HT y los niveles de TGF-β1, D1, D3 y GST-P en áreas focales/no focales. In vivo, HCB disminuyó los niveles de T3 y ARNm de la D1 y aumentó los niveles de T4 y ARNm de D3 del grupo DEN + HCB frente al grupo DEN. El HCB aumentó los niveles de D3, TGF-β1 y PCNA y disminuyó el D1 en las áreas focales. In vitro, HCB aumentó los niveles de PCNA, pSMAD 2/3 y TGF-β1 y la expresión de ARNm mientras que disminuyó los niveles de p21 y p27. El tratamiento con T3 exógeno previno las alteraciones moleculares relacionadas con la proliferación hepatocitaria. Estos efectos se evitaron utilizando un inhibidor del receptor II de TGF-β1. Los resultados sugieren que la alteración de la homeostasis de HT, a través de la D1 y la vía TGF-β1-SMAD, juega un papel clave en la proliferación celular y en las transformaciones neoplásicas tempranas en el HCC.

Animals , Rats , Carcinoma, Hepatocellular , Transforming Growth Factor beta1 , Iodide Peroxidase/genetics , Liver Neoplasms , Cell Proliferation
Arq. bras. endocrinol. metab ; 56(9): 614-617, Dec. 2012. ilus, tab
Article in English | LILACS | ID: lil-660275


OBJECTIVES: To investigate thyroid peroxidase gene (TPO) mutations in a Chinese siblings with congenital goitrous hypothyroidism (CGH). SUBJECTS AND METHODS: The proband, his sister, and their parents were enrolled. All subjects underwent clinical examination and laboratory tests. Mutation screening of the TPO gene was performed by sequencing fragments amplified from extracted genomic DNA. RESULTS: The siblings were diagnosed as CGH with neurodevelopmental deficits. Two compound heterozygous inactivating mutations were found in the two patients: a frameshift mutation between positions 2268 and 2269 (c.2268-2269 insT) and a missense mutation at c.2089 G>A (p.G667S) of the TPO gene. Their parents, with normal thyroid hormone levels, were heterozygous for mutations c.2268-2269 insT and c.2089 G>A, respectively. The polymorphisms of c.1207 G>T, c.1283 G>C, and c.2088 C>T were detected in the family. CONCLUSIONS: CGH of the Chinese siblings was due to the TPO gene mutations (c.2268-2269 insT and c.2089 G>A). Arq Bras Endocrinol Metab. 2012;56(9):614-7.

OBJETIVOS: Investigar mutações no gene da peroxidase da tireoide (TPO) em irmãos chineses com hipotireoidismo congênito com bócio (HCB). SUJEITOS E MÉTODOS: O probando, sua irmão e seus pais foram analisados. Todos os sujeitos passaram por exames clínicos e laboratoriais. A análise para mutações do gene TPO foi feita por meio de sequenciamento de fragmentos amplificados do DNA genômico extraído. RESULTADOS: Os irmãos foram diagnosticados com HCB e déficits de desenvolvimento neurológico. Duas mutações compostas, heterozigotas, inativadoras foram observadas nos dois pacientes: uma mutação frameshift entre as posições 2268 e 2269 (c.2268-2269 insT), e uma mutação missense em c.2089 G>A (p.G667S) do gene TPO. Os pais apresentaram níveis normais de hormônios da tiroide e eram heterozigotos para mutações em c.2268-2269 insT e c.2089 G>A, respectivamente. Foram detectados polimorfismos de c.1207 G>T, c.1283 G>C, e c.2088 C>T na família. CONCLUSÕES: O HCB dos irmãos chineses foi devido a mutações no gene TPO (c.2268-2269 insT e c.2089 G>A). Arq Bras Endocrinol Metab. 2012;56(9):614-7.

Adolescent , Female , Humans , Male , Young Adult , Congenital Hypothyroidism/genetics , Goiter/genetics , Iodide Peroxidase/genetics , Asian People/genetics , Frameshift Mutation/genetics , Mutation, Missense/genetics , Pedigree , Polymorphism, Genetic/genetics , Siblings
Arq. bras. endocrinol. metab ; 54(8): 732-737, Nov. 2010. ilus, tab
Article in English | LILACS | ID: lil-578348


The aim of this study was to identify the genetic defect of a patient with dyshormonogenetic congenital hypothyroidisms (CH) with total iodide organification defect (TIOD). A male child diagnosed with CH during neonatal screening. Laboratory tests confirmed the permanent and severe CH with TIOD (99 percent perchlorate release). The coding sequence of TPO, DUOX2, and DUOXA2 genes and 2957 base pairs (bp) of the TPO promoter were sequenced. Molecular analysis of patient's DNA identified the heterozygous duplication GGCC (c.1186_1187insGGCC) in exon 8 of the TPO gene. No additional mutation was detected either in the TPO gene, TPO promoter, DUOX2 or DUOXA2 genes. We have described a patient with a clear TIOD causing severe goitrous CH due to a monoallelic TPO mutation. A plausible explanation for the association between an autosomal recessive disorder with a single TPO-mutated allele is the presence of monoallelic TPO expression.

O objetivo deste estudo foi identificar defeitos genéticos em paciente com hipotireoidismo congênito (HC) por disormonogênese e defeito total de incorporação de iodeto (DIIT). Neonato do sexo masculino com HC diagnosticado pelo rastreamento neonatal. Exames clínicos e radiológicos confirmaram que o paciente apresentava HC severo e permanente com DIIT (teste de perclorato: 99 por cento). A região codificadora dos genes TPO, DUOX2, DUOXA2 e 2957 pares de bases (pb) do promotor de TPO foram sequenciados. No paciente foi identificada a duplicação em heterozigose GGCC no éxon 8 do gene TPO (c.1186_1187insGGCC). Nenhuma outra mutação foi localizada nos genes TPO, incluindo o promotor, DUOX2 ou DUOXA2. Descrevemos paciente com grave defeito de organificação de iodeto, provocando HC severo com bócio, em consequência de uma única mutação monoalélica no gene TPO. A expressão monoalélica no tecido tireoideano explicaria a associação de uma doen­ça autossômica recessiva com uma única mutação monoalélica.

Humans , Infant, Newborn , Male , Alleles , Autoantigens/genetics , Congenital Hypothyroidism/genetics , Iodide Peroxidase/genetics , Iron-Binding Proteins/genetics , Mutation/genetics , Sequence Analysis, DNA
Biol. Res ; 39(2): 307-319, 2006. ilus, tab
Article in English | LILACS | ID: lil-432433


Type-1 5'-iodothyronine deiodinase (5'-DI) is responsible for conversion of T4 to T3. Selenium (Se) is an integral part of this enzyme. Keeping in view the strong association between atherosclerosis and hypothyroidism, the present study examined the behavior of 5'-DI in liver, aorta and thyroid during hypercholesterolemia following different Se status, i.e., Se deficiency (0.02ppm), adequate (0.2ppm) and excess dose (1ppm) in SD male rats. Animals were fed a control or high-cholesterol diet (2%) for 1 and 2 months. 5'-DI activity and mRNA expression was measured by RIA and RT-PCR respectively. In liver and aorta, 5'-DI expression significantly decreased with the Se-deficient and the high-cholesterol diet. The trend was opposite in thyroid, i.e., mRNA expression increased significantly during selenium deficiency and with a high-cholesterol feeding. But with 1ppm Se supplementation, the 5'-DI expression increased in all the three tissues. The present study indicates that hypercholesterolemia along with selenium deficiency is co-responsible for differential regulation of 5'-DI enzyme in thyroidal vs. extrathyroidal tissues. Distinct regulation of 5'-DI in the thyroid reflects the clinical importance of this selenoprotein during hypercholesterolemia as this enzyme is essential for T3 production, which further has a vital role in the maintenance of lipid metabolism.

Animals , Male , Rats , Cholesterol, Dietary/administration & dosage , Hypercholesterolemia/metabolism , Iodide Peroxidase/metabolism , RNA, Messenger/metabolism , Selenium/analysis , Aorta/enzymology , Cholesterol, Dietary/metabolism , Hypercholesterolemia/enzymology , Iodide Peroxidase/genetics , Lipids/blood , Liver/enzymology , Radioimmunoassay , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Selenium/metabolism , Time Factors , Thyroid Gland/enzymology
Medicina (B.Aires) ; 65(3): 257-267, 2005. ilus
Article in Spanish | LILACS | ID: lil-425247


Las enfermedades tiroideas constituyen una heterogénea colección de anormalidades asociadas a mutaciones en los genes responsables en el desarrollo de la tiroides: factor de transcripción tiroideo 1 (TTF-1), factor de transcripción tiroideo 2 (TTF-2) y PAX8, o en uno de los genes que codifican para las proteínas involucradas en la biosíntesis de hormonas tiroideas como tiroglobulina (TG), tiroperoxidasa (TPO),sistema de generación de peróxido de hidrógeno (DUOX2), cotransportdor de Na/I– (NIS), pendrina (PDS), TSH y receptor de TSH. El hipotiroidismo congénito ocurre con una prevalencia de 1 en 4.000 nacidos. Los pacientes coneste síndrome pueden ser divididos en dos grupos: con hipotiroidismo congénito sin bocio (disembriogénesis) o con bocio (dishormonogénesis). El grupo de disembriogénesis, que corresponde al 85% de los casos, resulta de ectopía,agenesia o hipoplasia. En una minoría de estos pacientes, el hipotiroidismo congénito está asociado con mutaciones en los genes TTF-1, TTF-2, PAX-8, TSH o TSHr. La resencia de bocio congénito (15% de los casos) se ha asociado a mutaciones en los genes NIS, TG, TPO, DUOX2 o PDS. El hipotiroidismo congénito por dishormonogénesis es trasmitido en forma autonómica recesiva. Mutaciones somáticas en el TSHr han sido identificadas en adenomas tiroideos hiperfuncionantes. Otra enfermedad tiroidea bien establecida es la resistencia a hormonas tiroideas(RTH). Es un síndrome de reducida respuesta tisular a la acción hormonal causado por mutaciones localizadas en el gen del receptor de hormonas tiroideas (TR). Mutantes de TRinterfieren con la función del receptor normal por un mecanismo de dominancia negativa. En conclusión, la identificación de mutaciones en los genes de expresión tiroidea ha permitido un mayor entendimiento sobre la relación estructura-función de los mismos. La tiroides constituye un excelente modelo para el estudio molecular de las enfermedades genéticas.

Humans , Goiter/genetics , Hyperthyroidism/genetics , Hypothyroidism/genetics , Goiter/metabolism , Hyperthyroidism/metabolism , Hypothyroidism/congenital , Hypothyroidism/metabolism , Iodide Peroxidase/genetics , Iodide Peroxidase/metabolism , Mutation , Receptors, Thyrotropin/genetics , Thyroid Hormones/biosynthesis , Thyroid Hormones/genetics
Rev. Hosp. Clin. Fac. Med. Univ. Säo Paulo ; 45(1): 29-37, jan.-fev. 1990. ilus
Article in Portuguese | LILACS | ID: lil-97873


Os avanços da biologia molecular e o emprego de técnicas de identificaçäo de porçöes de DNA codificadoras para proteínas importantes na síntese, armazenamento, transporte e açäo periférica dos hormônios tireoideos, permitiram a elucidaçäo, a nivel molecular, da etiologia de várias condiçöes genéticas da tireóide. Em pacientes com defeito na peroxidase tireóidea (TPO) observou-se a presença de polimorfismo (RFLP) com a endonuclease Bgl-I que se segrega, em 2 alelos, com o fenótipo com bócio, hipotireoidismo e teste de perclorato positivo. Em defeitos da tireoglobulina (Tg), observou-se nível quantitativamente baixo de Tg, concomitante com a virtual ausência da RNA mensageiro para a Ig. Por outro lado, a ausência de incorporaçäo de ácido siálico na molécula de Tg, produz proteína com defeito estrutural que impede a síntese adequada de T3 e T4. Tais defeitos da Tg säo igualmente encontrados em animais com bócio congênito (gados bovino e caprino, e camundongos cog/cog). A presença de mutaçäo puntiforme no nucleotídeo 281 do exon 5 do RNA da globulina carreadora de T4 (TBG) leva a expressäo genética de proteína instável, com queda da TBG sérica. O efeito nos receptores nucleares de T3 estäo ligados ao gene codificador (c-erb-A-beta) localizado no cormossoma 3. A presença de polimorfismo EcorRV nos indivíduos, sugere gene mutante codificando proteína que poderia näo fixar T3 ou produzir proteína anômala com atividade bloqueadora da açäo de T3 impedindo a fixaçäo nuclear

Humans , Animals , Thyroid Hormones/genetics , Molecular Biology , Cloning, Molecular , DNA, Recombinant/genetics , DNA, Recombinant/metabolism , Genomic Library , Hypothyroidism/genetics , Hypothyroidism/metabolism , Iodide Peroxidase/genetics , Iodide Peroxidase/metabolism , Receptors, Thyroid Hormone/genetics , Thyroglobulin/genetics , Thyroglobulin/metabolism