Foxp3 expression is associated with aggressiveness in differentiated thyroid carcinomas

OBJECTIVES: Forkhead box P3 (FoxP3) expression has been observed in human cancer cells but has not yet been reported in thyroid cells. We investigated the prognostic significance of both FoxP3 expression and intratumoral FoxP3+ lymphocyte infiltration in differentiated thyroid carcinoma cells. METHODS: We constructed a tissue microarray with 385 thyroid tissues, including 266 malignant tissues (from 253 papillary thyroid carcinomas and 13 follicular carcinomas), 114 benign lesions, and 5 normal thyroid tissues. RESULTS: We determined the expression of FoxP3 in both tumor cells and tumor-infiltrating lymphocytes using immunohistochemical techniques. Cellular expression of FoxP3 was evident in 71% of benign and 91.9% of malignant tissues. The nuclear and cytoplasmic expression patterns were quantified separately. A multivariate logistic regression analysis indicated that cytoplasmic FoxP3 expression is an independent risk factor for thyroid malignancy. Cytoplasmic FoxP3 staining was inversely correlated with patient age. Nuclear FoxP3 staining was more intense in younger patients and in tumors presenting with metastasis at diagnosis. FoxP3+ lymphocytes were more frequent in tumors smaller than 2 cm, those without extrathyroidal invasion, and in patients with concurrent chronic lymphocytic thyroiditis. CONCLUSIONS: We demonstrated FoxP3 expression in differentiated thyroid carcinoma cells and found evidence that this expression may exert an important influence on several features of tumor aggressiveness

P53 And Expression Of Immunological Markers May Identify Early Stage Thyroid Tumors.

Besides its major role in cell proliferation, DNA repair, and apoptosis, functional p53 protein is involved in the induction of antitumor cytotoxic-T-cell activity against carcinoma cells. We aimed to investigate p53 and immune cell markers utility as diagnostic and prognostic markers of differentiated thyroid cancer (DTC). ACIS-III system was used to evaluate p53 and immune cell markers including tumor-associated macrophages (TAM); CD68 and tumor-infiltrating lymphocytes (TIL) subsets such as CD3, CD4, CD8, and CD20 in 206 thyroid carcinomas, 105 benign nodules, and 18 normal tissues. Also, TP53 was sequenced in 78 out of 164 patients with papillary thyroid carcinoma. P53 expression was observed more frequently in malignant than in benign lesions (P < 0.0001) and helped discriminate follicular patterned lesions. In addition, p53 was more frequent in smaller (P = 0.0015), unique tumors (P = 0.0286), with thyroiditis (P = 0.0486) and without metastasis at diagnosis (P = 0.0201). TAM was more frequent in P53 negative tumors (P = 0.002). Infiltration of CD8+ TIL was found in 61.7% of P53 positive and 25.6% of P53 negative DTC (P < 0.001). We suggest that p53 and CD8+ TIL immune profile analysis might be useful in DTC.201384658

Identifying A Risk Profile For Thyroid Cancer.

The large use of simple and effective diagnostic tools has significantly contributed to the increase in diagnosis of thyroid cancer over the past years. However, there is compelling evidence that most micropapillary carcinomas have an indolent behavior and may never evolve into clinical cancers. Therefore, there is an urgent need for new tools able to predict which thyroid cancers will remain silent, and which thyroid cancers will present an aggressive behavior. There are a number of well-established clinical predictors of malignancy and recent studies have suggested that some of the patients laboratory data and image methods may be useful. Molecular markers have also been increasingly tested and some of them appear to be very promising, such as BRAF, a few GST genes and p53 polymorphisms. In addition, modern tools, such as immunocytochemical markers, and the measure of the fractal nature of chromatin organization may increase the specificity of the pathological diagnosis of malignancy and help ascertain the prognosis. Guidelines designed to select nodules for further evaluation, as well as new methods aimed at distinguishing carcinomas of higher aggressiveness among the usually indolent thyroid tumors are an utmost necessity.51713-2

Evaluation of prognostic of the thyroid carcinoma through immunohistochemical markers

Orientador: Laura Sterian WardTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias MedicasResumo: Como a maior parte dos carcinomas diferenciados tem um excelente prognóstico, alguns autores consideram o tratamento convencional, isto é, a tireoidectomia total seguida de ablação actínica de remanescentes glandulares, excessivo. Combinações de fatores específicos do paciente e do tumor foram propostas para classificação prognóstica, mas na prática são pouco utilizadas. Os resultados de alguns trabalhos sugerem relação entre a expressão dos genes MUC, p53 e NIS e a evolução clínica das neoplasias tireoidianas. Com o objetivo de investigar a utilidade clínica de marcadores imonoistoquímicos de prognóstico, estudamos 67 nódulos tireoidianos, incluindo 57 carcinomas, dos quais incluiu 34 (50,74%) carcinomas papilíferos, sendo 21 (31,3%) casos do tipo histológico clássico, 7 (10,4%) da variante de células altas e 6 (9%) da variante folicular. Avaliamos também 16 (23,9%) carcinomas foliculares, 4 (6%) medulares e 3 carcinomas anaplásicos (4,5%). Dez casos (14,9%) foram obtidos de pacientes submetidos à cirurgia com o diagnóstico de doenças benignas da tiróide (3 casos de bócio nodular, 4 adenomas foliculares e 3 tecidos tireoidianos normais). Todos os pacientes foram submetidos a um mesmo protocolo de tratamento e acompanhados por 66±28 meses (mediana de 63 meses) sendo classificados como apresentando evolução favorável ou desfavorável. Consideramos de evolução desfavorável os 19 pacientes que possuíam evidência de recorrência local ou à distância ou que foram a óbito durante o seguimento pela doença. Realizamos estudos imunoistoquímicos da detecção das proteínas NIS, p53 e MUC1 comparando-os com a evolução dos pacientes. Mostramos que pacientes com carcinomas bem diferenciados, ausência de metástases, estadio II e estadiamento T2 têm uma evolução melhor em relação aos pacientes com carcinomas pouco diferenciados, presença de metástases, estadio IV e estadiamento T4. Não encontramos relação entre a expressão das proteínas NIS, p53 e MUC1 e a evolução dos pacientes estudadosAbstract: Because most differentiated thyroid carcinomas have an excellent prognosis, some authors have been claiming that these patients have been over-treated. Combinations of patient-and tumor-specific factors have been proposed for prognostic stratification, but no clinicpathologic staging was demonstrated to be useful at the present time. In order to investigate the clinical utility of immunohistochemistry markers, we studied 67 thyroid nodules including 50 thyroid carcinomas. There were 34 (50,74%) papillary carcinomas, including 21 (31,3%) cases of the classical histological type, 7 (10,4%) tall cell variants and 6 (9%) of the follicular variant type. Also, there were 16 (23,9%) follicular carcinomas, 4 (6%) medullary and 3 anaplastic carcinomas (4,5%). Ten patients (14,9%) were submitted to surgery because of benign thyroid diseases (3 nodular goiter cases, 4 follicular adenomas and 3 thyroid tissues normal). All patients were submitted to a similar management protocol and followed-up for 66±28 months (median of 63 months) and classified as presenting a good or a bad evolution. We considered as presenting a bad evolution 19 patients that presented evidence of local or distant recurrence and the patients that died during the follow-up. Immunohistochemical expression of NIS, p53 and MUC1 proteins was compared to patient¿s evolution. We showed that patient with well differentiated thyroid carcinomas, absence of distant metastases, stage II and staging T2 have a better evolution in relation to the patients with poorly differentiated thyroid carcinomas, presence of metastases, stage IV and staging T4. We were able to find relation between the expression of proteins NIS, p53 and MUC1 and prognostic of the studied patientsDoutoradoClinica MedicaDoutor em Clínica Médic

Estudo do sistema glutationa S-transferase nos tumores da tiroide humana

Orientador : Laura Sterian WardDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: A susceptibilidade a carcinógenos químicos tem um importante papel no desenvolvimento da maioria dos cânceres. Diversos polimorfismos de enzimas metabolizadoras de drogas em humanos influenciam esta susceptibilidade individual. Os genes que incluem as isoenzimas do sistema glutationa s-transferase (GST) apresentam um polimorfismo hereditário. Os genes GST mu 1 (GSTM1) and GST theta 1(GSTT1) possuem uma variante alélica nula nos quais o gene inteiro está ausente e, portanto, não existe codificação ou produção da respectiva enzima. O genótipo nulo para ambas enzimas foi associado com diferentes tipos de cânceres. Para observar a influência da herança dessas enzimas no risco do câncer da tiróide, utilizamos uma multiplex-PCR que inclui o gene p-globina como um controle de DNA na reação de PCR para comparar 300 indivíduos normais de nossa população com 116 pacientes portadores de lesões tiroidianas. Desses casos, 49 eram de bócio benigno e 67 de doenças malignas: 50 carcinomas papilíferos e 17 carcinomas foliculares. A comparação entre tecidos de tumores da tiróide e 35 amostras correspondentes de sangue periférico de pacientes com câncer demonstraram padrão idêntico, sugerindo que o sistema GST não está envolvido no processo de desdiferenciação folicular. Não houve diferença estatística entre a prevalência da deleção dos genes GSTT1 e GSTM1 em indivíduos normais e em pacientes com bócio. Entretanto, os pacientes com carcinoma papilífero (10%) e pacientes com carcinoma folicular (17%) apresentaram uma prevalência de genótipo nulo mais elevada do que os indivíduos da população normal (5%) (p= 0,0479). Demonstramos que a herança de um genótipo nulo combinado para GSTM1 e GSTT1 é responsável por um aumento de 2.6 vezes no risco de câncer. Sugerimos que estes resultados podem ser usados como marcadores na susceptibilidade ao câncer da tiróide, auxiliando na seleção dos indivíduos de risco que merecem investigação e conduta mais rigorosa de seus bóciosAbstract: Susceptibility to chemical carcinogens plays an important role in the development of most cancers. Several polymorphisms of human drug-metabolizing enzymes influence this individual susceptibility. The genes that encode the isoenzymes of the glutathione s-transferase (GST) system present a polymorphic inheritance. The GST mu 1 (GSTMl) and GST theta 1(GSTTl) genes have a null allele variant in which the entire gene is absent. The null genotype for both enzymes has been associated with many different types of tumors. In order to look for the influence ofthe inheritance pattern of these enzymes on thyroid cancer risk we used a triplex PCRi that included p-globin gene as a DNA quality control in the PCR reaction to compare 300 normal individuaIs of our population to 116 goiter patients. There were 49 cases of benign goiters and 67 cases of malignant diseases: 50 papillary and 17 follicular carcinomas. Comparison between thyroid tumor specimens and normal corresponding samples of 35 cancer patients demonstrated identical patterns, suggesting that the GST system is not involved in the process of follicular dedifferentiation. There was no statistical difference between the prevalence ofthe deleted alleles in the normal individuaIs and in the goiter patients. However, papillary carcinoma patients (10%) and follieular carcinoma patients (17%) presented a higher prevaIenee of the null genotype than the normal population individuaIs (5%) (p= 0,0479). We found a 2.6 increased risk of thyroid caneer associated with the GSTT1 and GSTMl combined null inheritance, suggesting that this may be a usefuI marker for thyroid caneer susceptibilityMestradoCiencias BasicasMestre em Clinica Medic

Cd8+ Tumour-infiltrating Lymphocytes And Cox2 Expression May Predict Relapse In Differentiated Thyroid Cancer.

There is an increasing rate of papillary thyroid carcinomas that may never progress to cause symptoms or death. Predicting outcome and determining tumour aggressiveness could help diminish the number of patients submitted to aggressive treatments. We aimed to evaluate whether markers of the immune system response and of tumour-associated inflammation could predict outcome of differentiated thyroid cancer (DTC) patients. Retrospective cohort study. We studied 399 consecutive patients, including 325 papillary and 74 follicular thyroid carcinomas. Immune cell markers were evaluated using immunohistochemistry, including tumour-associated macrophages (CD68) and subsets of tumour-infiltrating lymphocytes (TIL), such as CD3, CD4, CD8, CD16, CD20, CD45RO, GRANZYME B, CD69 and CD25. We also investigated the expression of cyclooxygenase 2 (COX2) in tumour cells and the presence of concurrent lymphocytic infiltration characterizing chronic thyroiditis. Concurrent lymphocytic infiltration characterizing chronic thyroiditis was observed in 29% of the cases. Among all the immunological parameters evaluated, only the enrichment of CD8+ lymphocytes (P = 0·001) and expression of COX2 (P =0·01) were associated with recurrence. A multivariate model analysis identified CD8+ TIL/COX2 as independent risk factor for recurrence. A multivariate analysis using Cox's proportional-hazards model adjusted for the presence of concurrent chronic thyroiditis demonstrated that the presence of concurrent chronic thyroiditis had no effect on prognostic prediction mediated by CD8+ TIL and COX2. In conclusion, we suggest the use of a relatively simple pathology tool to help select cases that may benefit of a more aggressive approach sparing the majority of patients from unnecessary procedures

Lack Of Mutation In Exon 10 Of P53 Gene In Thyroid Tumors.

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CD8+ tumour‐infiltrating lymphocytes and COX2 expression may predict relapse in differentiated thyroid cancer

There is an increasing rate of papillary thyroid carcinomas that may never progress to cause symptoms or death. Predicting outcome and determining tumour aggressiveness could help diminish the number of patients submitted to aggressive treatments. We aimed to evaluate whether markers of the immune system response and of tumour‐associated inflammation could predict outcome of differentiated thyroid cancer (DTC) patients. Retrospective cohort study. We studied 399 consecutive patients, including 325 papillary and 74 follicular thyroid carcinomas. Immune cell markers were evaluated using immunohistochemistry, including tumour‐associated macrophages (CD68) and subsets of tumour‐infiltrating lymphocytes (TIL), such as CD3, CD4, CD8, CD16, CD20, CD45RO, GRANZYME B, CD69 and CD25. We also investigated the expression of cyclooxygenase 2 (COX2) in tumour cells and the presence of concurrent lymphocytic infiltration characterizing chronic thyroiditis. Concurrent lymphocytic infiltration characterizing chronic thyroiditis was observed in 29% of the cases. Among all the immunological parameters evaluated, only the enrichment of CD8+ lymphocytes (P = 0·001) and expression of COX2 (P =0·01) were associated with recurrence. A multivariate model analysis identified CD8+ TIL/COX2 as independent risk factor for recurrence. A multivariate analysis using Cox's proportional‐hazards model adjusted for the presence of concurrent chronic thyroiditis demonstrated that the presence of concurrent chronic thyroiditis had no effect on prognostic prediction mediated by CD8+ TIL and COX2. In conclusion, we suggest the use of a relatively simple pathology tool to help select cases that may benefit of a more aggressive approach sparing the majority of patients from unnecessary procedures.832246253FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2010/00394-0; 2011/19612-

mRNA BRAF expression helps to identify papillary thyroid carcinomas in thyroid nodules independently of the presence of BRAFV600E mutation

Literature has consistently shown associations of BRAFV600E mutation with papillary thyroid cancer clinical features. However, the clinical utility of BRAF expression has not been clinically explored so far. We studied 67 thyroid nodules (32 benign nodules and 35 PTC cases). BRAF mRNA expression levels measured by a quantitative real-time PCR and a PCR-RFLP were used to identify BRAFV600E mutation. BRAF mRNA expression was significantly higher in malignant (198.2 +/- 373.9 AU) than in benign (4.1 +/- 6.9 AU) nodules (p < 0.0001). BRAF expression identified malignancy with a sensitivity of 80.6%, specificity of 77.1%, positive predictive value of 75.8%, and negative predictive value of 81.8%. A cut-point of 4.712, identified by the ROC curve, was able to sort out malignant nodules with an accuracy of 78.8%. Although we did not find any correlation between the presence of BRAF V600E mutation and clinical or tumor features such as age (p = 0.309), gender (p = 0.5453), ethnicity (p = 0.9820), tumor size (p = 1.000), multifocality (p = 0.2530) or mRNA levels (p = 0.7510), the study power for BRAF expression and diagnosis (99%; FPRP = 0.85) indicated that data is noteworthy despite the relative small number of patients investigated. We concluded that BRAF mRNA expression may help to identify PTC among thyroid nodules independently of the presence of BRAFV600E mutation. (C) 2012 Elsevier GmbH. All rights reserved.208848949