Neither bovine somatotropin nor growth hormone-releasing factor alters expression of thyroid hormone receptors in liver and mammary tissues

Physiological effects of thyroid hormones are mediated primarily by binding of triiodothyronine to specific nuclear receptors. Organ-specific changes in production of triiodothyronine from its prohormone, thyroxine, have been hypothesized to target the action of thyroid hormones on the mammary gland and play a role in mediating or augmenting a galactopoietic response to bovine somatotropin (bST). Additionally, tissue responsiveness to thyroid hormones may be altered by changes in the number or affinity of nuclear receptors for thyroid hormones. In the present study, effects of bST and bovine growth hormone-releasing factor (bGRF) on thyroid hormone receptors in liver and mammary gland were studied. Lactating Holstein cows received continuous infusions of bST or bGRF for 63 d or served as uninfused controls. Nuclei were isolated from harvested mammary and liver tissues and incubated with [(125)I]-triiodothyronine. Treatments did not alter the capacity or affinity of specific binding sites for triiodothyronine in liver or mammary nuclei. Evaluation of transcript abundance for thyroid hormone receptors showed that isoforms of thyroid hormone receptor or retinoid receptor (which may influence thyroid receptor action) expressed in the mammary gland were not altered by bST or bGRF treatment. Data do not support the hypothesis that administration of bST or bGRF alters sensitivity of mammary tissue by changing expression of thyroid hormone receptors.

The 434(G > C) polymorphism in the eosinophil cationic protein gene and its association with tissue eosinophilia in oral squamous cell carcinomas

Objective: The aim of this study was to investigate the prevalence of the Eosinophil cationic protein (ECP)-gene polymorphism 434(G > C) in oral squamous cell carcinoma (OSCC) patients and its association with tumor-associated tissue eosinophilia (TATE), demographic, clinical, and microscopic variables. Methods: The ECP genotypes of 165 healthy individuals and 157 OSCC patients were detected by PCR-RFLP analysis after cleavage of the amplified DNA sequence with enzyme PstI. TATE was obtained by morphometric analysis. Chi-square test or Fisher`s exact test was used to analyze the association of ECP-gene polymorphism 434(G > C) with TATE, demographic, clinical, and microscopic variables in OSCC patients. Disease-free survival and overall survival were calculated by the Kaplan-Meier product-limit actuarial method and the comparison of the survival curves were performed using log rank test. Results: Most of healthy individuals (53.33%) and OSCC patients (57.97%) were heterozygous for the ECP 434(G > C) polymorphism. Based on numerical differences, our results showed that OSCC patients with intense TATE and at least one C allele had a higher frequency of bilateral neck dissection, local recurrence, vascular embolization, involved resection margins, and postoperative radiotherapy. No statistically significant differences on survival rates were found in OSCC patients presenting different ECP 434(G > C) genotypes. Conclusions: These results suggest a tendency towards a poor clinical outcome in OSCC patients with intense TATE and 434GC/CC genotypes, probably due to an ECP genetic variant with altered cytotoxic activity.

Interrelationship between Epstein-Barr virus infection in gastric carcinomas and the expression of apoptosis-associated proteins

Aims: Epstein-Barr virus (EBV) and its associated proteins may be protective against the occurrence of apoptosis that would normally inhibit cancer development and progression. Alternatively, the viral infection may cause altered or mutated expression of oncogenes or tumour suppressor genes that are necessary for tumour development. an action that may also involve apoptosis, In this study, a relationship was sought between occurrence of EBV infection, expression of apoptosis-associated proteins (tumour suppressor gene p53 and oncogenes c-myc and bcl-2) and levels of cell death (apoptosis or necrosis) in 119 cases of gastric carcinoma. Methods and results: The EBV status of the gastric carcinomas (using the EBV-encoded small RNA I (EBER-1) and in-situ hybridization), stage and grade of tumour and sex of patients were compared for bcl-2, p53 and c-myc expression patterns. EBER-1 was detected in approximately 20% of cases studied. There was no significant correlation between levels of cell death in the tumour tissue and EBV status. In the protein analyses, development and progression of gastric carcinoma, with or without EBV infection. was independent of bcl-2 expression. However, in gastric cancers with EBV infection, p53 overexpression was inhibited and c-myc expression was increased in early stage cancers, in comparison with decreased c-myc expression in late stage cancers. Conclusions: The p53 and c-myc expression patterns indicate that EBV-infected gastric carcinomas are less likely to have a natural regression via apoptosis at an early stage and explain, in part, the resistance to treatment of late stage of gastric cancers.

Apoptosis and Expression of Bcl-2, Bcl-XL, and Bax in Renal Cell Carcinomas

There are at present disparate published results with regard to the relevance of the Bcl-2 gene family, levels of apoptosis, and cell proliferation in the development and progression of renal cell carcinoma (RCC). The present study v analyses the interrelationship between the expression of representatives of the anti-apoptotic (Bcl-2, Bcl-X-L) or pro-apoptotic (Bax) Bcl-2 proteins, incidence of apoptosis, and mitosis in a selected small group of 22 graded RCCs that had paired normal renal tissue, or non-neoplastic tissue in the renal biopsy specimen. The cases were chosen to determine the feasibility of measuring these parameters as potential surrogate markers of progression or treatment failure of the cancers. The results showed that in approximately 50% of the RCCs, where Bcl-2 and/or Bcl-X-L expression was high, apoptosis it-as not detected, and when expression of these proteins was low or not found, increased levels of apoptosis were seen. In most of the remaining 50% of samples, high levels of Bcl-X-L but not Bcl-2 were negatively correlated with low levels of apoptosis (Bcl-X-L: r = -0.437, P = 0.07 and Bcl-2: r = + 0.560, P = 0.02). For the same group of samples, high Bax expression was found in association with apoptosis (r = + 0.578, P = 0,02). A novel finding was an association between low expression of Bcl-2 an/or Bcl-X-L in normal tissue and the level of expression of these proteins in the RCCs, an intrinsic variation that may be an individual patient factor. The results indicate that, in RCCs with increased expression of Bcl-2 and/or Bcl-X-L, levels of apoptosis are minimal and these combined factors may assist in progression of the cancers and resistance to treatments.

Expression of a truncated Brca1 protein delays lactational mammary development in transgenic mice

To address the hypothesis that certain disease-associated mutants of the breast-ovarian cancer susceptibility gene BRCA1 have biological activity in vivo, we have expressed a truncated Brca1 protein (trBrca1) in cell-lines and in the mammary gland of transgenic mice. Immunofluorescent analysis of transfected cell-lines indicates that trBRCA1 is a stable protein and that it is localized in the cell cytoplasm. Functional analysis of these cell-lines indicates that expression of trBRCA1 confers an increased radiosensitivity phenotype on mammary epithelial cells, consistent with abrogation of the BRCA1 pathway. MMTV-trBrca1 transgenic mice from two independent lines displayed a delay in lactational mammary gland development, as demonstrated by altered histological profiles of lobuloalveolar structures. Cellular and molecular analyses indicate that this phenotype results from a defect in differentiation, rather than altered rates of proliferation or apoptosis. The results presented in this paper are consistent with trBrca1 possessing dominant-negative activity and playing an important role in regulating normal mammary development. They may also have implications for germline carriers of BRCA1 mutations.

Apoptosis and cell proliferation in the development of gastric carcinomas: Associations with c-myc and p53 protein expression

Background and Aim: Patients with gastric carcinomas have a poor prognosis and low survival rates. The aim of the present paper was to characterize cellular and molecular properties to provide insight into aspects of tumor progression in early compared with advanced gastric cancers. Methods: One hundred and nine graded gastric carcinomas (early or advanced stage, undifferentiated or differentiated type) with paired non-cancer tissue were studied to define the correlation between apoptosis (morphology, terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling), cell proliferation (Ki-67 expression, morphology) and expression and localization of two proteins frequently having altered expression in cancers, namely p53 and c-myc. Results: Overall, apoptosis was lower in early stage, differentiated and undifferentiated gastric carcinomas compared with advanced-stage cancers. Cell proliferation was comparatively high in all stages. There was a high level of p53 positivity in all stages. Only the early- and advanced-stage undifferentiated cancers that were p53 positive had a significantly higher level of apoptosis (P< 0.05). Cell proliferation was significantly greater (P < 0.05) only in the early undifferentiated cancers that had either c-myc or p53-positivity. Conclusions: The results indicate that low apoptosis and high cell proliferation combine to drive gastric cancer development. The molecular controls for high cell proliferation of the early stage undifferentiated gastric cancers involve overexpression of both p53 and c-myc. Overexpression of p53 may also control cancer development in that its expression is associated with higher levels of apoptosis in early and late-stage undifferentiated, cancers. (C) 2002 Blackwell Publishing Asia Pty Ltd.

Reciprocal relationship between methylation status and loss of heterozygosity at the p14 (ARF) locus in Australian and South African hepatocellular carcinomas

Chromosome 9p21, a locus comprising the tumor suppressor genes (TSG) p16(INK4) (a) and p14(ARF) , is a common region of loss of heterozygosity (LOH) in hepatocellular carcinoma (HCC). p14(ARF) shares exon 2 with p16 in a different reading frame. p14 binds to MDM2 resulting in a stabilization of functional p53 . This study examined the roles of p14, p16 and p53 in hepatocarcinogenesis, in 37 Australian and 24 South African patients. LOH at 9p21 and 17p13.1, p14 and p16 mutation analysis, p14 and p16 promoter methylation and p14, p16 and p53 protein expression was examined. LOH at 9p21 was detected more frequently in South African HCC (P = 0.04). Comparable rates of p53 LOH were observed in Australian and South African HCC (10/22, 45%vs 13/22, 59%, respectively). Hypermethylation of the p14 promoter was more prevalent in Australian HCC than in South African HCC (17/37, 46%vs 7/24, 29%, respectively). In Australian HCC the prevalence of p14 methylation increased with age (P = 0.03). p16 promoter methylation was observed in 12/37 (32%) and 6/24 (25%) in Australian and South African HCC, respectively. Loss of p16 protein expression was detected in 14/36 Australian HCC whereas p53 protein expression was detected in 9/36. Significantly, a reciprocal relationship between 9p21 LOH and p14 promoter hypermethylation was observed (P less than or equal to0.05 ). No significant association between p14 and p53 was seen in this study. The reciprocal relationship identified indicates different pathways of tumorigenesis and likely reflects different etiologies of HCC in the two countries. (C) 2002 Blackwell Science Asia Pty Ltd.

Peroxisome proliferator-activated receptor alpha in the human breast cancer cell lines MCF-7 and MDA-MB-231

Peroxisome proliferator-activated receptor (PPAR) alpha is a ligand-activated transcription factor that has been linked with rodent hepatocarcinogenesis. It has been suggested that PPARalpha mRNA expression levels are an important determinant of rodent hepatic tumorigenicity. Previous work in rat mammary gland epithelial cells showed significantly increased PPARalpha mRNA expression in carcinomas, suggesting the possible role of this isoform in rodent mammary gland carcinogenesis. In this study we sought to determine whether PPARalpha is expressed and dynamically regulated in human breast cancer MCF-7 and MDA-MB-231 cells. Having established the presence of PPARalpha in both cell types, we then examined the consequence of PPARa activation, by its ligands Wy-14,643 and clofibrate, on proliferation. With real-time reverse transcriptase-polymerase chain reaction, we showed that PPARalpha mRNA was dynamically regulated in MDA-MB-231 cells and that PPARalpha activation significantly increased proliferation of the cell line. In contrast, PPARalpha expression in MCF-7 cells did not change with proliferation during culture and was present at significantly lower levels than in MDA-MB-231 cells. However, PPARalpha ligand activation still significantly increased the proliferation of MCF-7 cells. The promotion of proliferation in breast cancer cell lines following PPARalpha activation was in stark contrast to the effects of PPARgamma-activating ligands that decrease proliferation in human breast cancer cells. our results established the presence of PPARalpha in human breast cancer cell lines and showed for the first time that activation of PPARalpha in human breast cancer cells promoted proliferation. Hence, this pathway may be significant in mammary gland tumorigenesis. (C) 2002 Wiley-Liss, Inc.

O papel das proteínas da matriz extracelular e das metaloproteinases em carcinomas de cabeça e pescoço: uma atualização bibliográfica

Interações entre células neoplásicas e constituintes da matriz extracelular (MEC) interferem fortemente no desenvolvimento tumoral, incluindo os localizados em cabeça e pescoço, pois influenciam a proliferação e sobrevivência celular, bem como a sua capacidade de migrar do sítio primário para outros tecidos e formar metástases. Essa migração celular é facilitada pela destruição parcial da MEC, a qual é realizada pelas metaloproteinases (MMPs), que representam uma família de mais de vinte endopeptidases, com atividade controlada pela expressão de inibidores específicos (TIMPs). Diversos estudos utilizando-se de marcadores para constituintes da MEC bem como pelas MMPs têm fornecido informações adicionais sobre o diagnóstico e prognóstico em carcinomas de cabeça e pescoço. Nesta revisão consideraremos o papel da MEC e das MMPs na progressão desses tumores, enfatizando que não somente a degradação proteolítica está envolvida neste processo, como também interações entre vários constituintes da MEC fornecem substrato para regulação e crescimento destes tumores.

Avaliação de fatores demográficos e estadiamento clínico nos pacientes submetidos a resgate cirúrgico por carcinomas epidermóides de orofaringe

O tratamento clássico do carcinoma epidermóide das vias aéreas digestivas superiores (VADS) é a cirurgia, associada ou não à radioterapia pós-operatória. As recidivas loco-regionais constituem a principal falha no tratamento inicial e o diagnóstico precoce favorece a indicação de cirurgia de resgate. OBJETIVO: Análise descritiva dos dados demográficos e de estadiamento no resgate cirúrgico dos tumores de orofaringe. CASUÍSTICO E MÉTODOS: Estudo retrospectivo de 78 pacientes, submetidos à cirurgia em todos os casos e a radioterapia complementar em 37 destes. RESULTADOS: Predomínio do gênero masculino em 70 casos, com idade média de 54,2 anos e etnia branca em 54 pacientes. O pacientes foram estadiados em T3 e T4 em 38 casos. Dos 78 pacientes, 40 eram pescoço N0. Trinta e cinco pacientes desenvolveram recidiva loco-regional e à distância. Das recidivas loco-regionais, 17 deles foram submetidos ao resgate cirúrgico, sendo que 12 pacientes foram reestadiados na recidiva em T1; 2 em T2 e quanto ao N, 2 eram N2a e 2 eram N2b. A idade média dos pacientes submetidos ao resgate foi de 52,8 anos, com predomínio na etnia branca e gênero masculino. CONCLUSÃO: O resgate predominou em estádios I e II com controle da doença em 58,8%.

Valor prognóstico da correlação imunoistoquímica do Ki-67 e p53 em carcinomas epidermóides da laringe

Os fatores prognósticos histológicos podem colaborar para determinar a evolução desta neoplasia. OBJETIVO: Correlacionar a expressão imunoistoquímica do p53 e Ki-67 com idade, grau histológico, comprometimento de linfonodos e estadiamento patológico de pacientes com carcinomas epidermóides de laringe. MÉTODO: Foram avaliados trinta casos consecutivos de carcinomas epidermóides de laringe, submetidos à técnica de imunoistoquímica para verificar a expressão dos anticorpos p53 e Ki-67. RESULTADO: A idade média foi de 56,2 anos e a imunoexpressão dos marcadores foi mais observada no grupo com mais de 50 anos de idade, especialmente a do anticorpo Ki-67 (p=0,032). Não houve relação do p53 e Ki-67 com o comprometimento de linfonodos. O Ki-67 se expressou em 70% nos casos de alto grau histológico e 80% nos de baixo, enquanto o p53 em 70% apenas nos de alto grau. O estadiamento patológico mostrou que no grupo dos carcinomas avançados a expressão do p53 foi de 61,5%, enquanto o Ki-67 mostrou positividade para os casos precoces (100%) e avançados (73,1%). CONCLUSÃO: Não houve diferenças significantes entre a imunoexpressão do p53 e do Ki-67 em carcinomas epidermóides de laringe, exceto no grupo de pacientes com mais de 50 anos, onde a expressão do Ki-67 foi significativamente maior.

Quantificação da difusão na ressonância magnética da mama: ADC e Kurtosis

Mestrado em Radiações Aplicadas às Tecnologias da Saúde. Área de especialização: Ressonância Magnética

Quantificação por imagem ponderada em difusão (DWI) das lesões tumorais da mama

Introdução – A técnica de Difusão por Ressonância Magnética (RM), ao avaliar o movimento das moléculas de água nos tecidos, permite inferir sobre a arquitetura dos mesmos, em particular relativamente à celularidade, volume celular e permeabilidade das membranas. O Coeficiente de Difusão Aparente (ADC) é um parâmetro quantificável da imagem ponderada em difusão (DWI). A sua análise poderá fornecer informação clínica adicional sobre estas lesões, sobretudo em relação à sua caracterização histológica. Objetivos – Caracterizar e diferenciar tipos e alguns subtipos de lesões mamárias através da análise do ADC. Metodologia – 20 Mulheres com 23 lesões mamárias foram submetidas a RM mamária: 3 lesões benignas (3 Fibroadenomas-FA) e 20 malignas (16 Carcinomas Ductais Invasivos-CDI, 2 Carcinomas Ductais In Situ-CDIS e 2 Carcinomas Invasivos Lobulares-CLI). Num equipamento 1.5T aplicou-se uma sequência de Difusão (b=0,50,250,500,750,1000 s/mm2). Obteve-se o ADC através do ajuste exponencial da intensidade de sinal das lesões em função do valor de b, fazendo-se corresponder os valores de ADC à respetiva caracterização histológica e posterior comparação com a literatura. Resultados e Discussão – As lesões malignas apresentaram ADCs significativamente (p=0,014) inferiores [(0,94±0,22)x10-3 mm2/s] aos das benignas [(1,43±0,25)x10-3 mm2/s]. A justificação pode residir no aumento da celularidade e consequente restrição da Difusão que se observa nas lesões malignas. Os CDI apresentaram ADCs baixos [(0,88±0,17)x10-3 mm2/s], enquanto que os CDIS apresentaram ADCs mais elevados [(1,33±0,29)x10-3 mm2/s]. Estes resultados estão de acordo com o facto dos CDIS estarem limitados aos ductos mamários, mantendo-se menos alterada a estrutura do tecido adjacente e resultando numa menor restrição à difusão que nos CDI. Verificaram-se diferenças significativas entre FA e CDI (p=0,010) e entre CDI e CDIS (p=0,049). Conclusões – O ADC possibilita a diferenciação entre lesões mamárias benignas e malignas, bem como entre alguns tipos histológicos. O desenvolvimento deste conceito pode representar um avanço no papel da RM na avaliação destas neoplasias. ABSTRACT - Introduction – The Magnetic Resonance (MR) diffusion technique measures the movement of water molecules in tissues. Therefore, it provides useful information about tissue architecture, specially regarding tissue cellularity, cell volume and membrane permeability. The quantification of diffusion weighted imaging (DWI) data is done by measuring the so-called. Apparent Diffusion Coefficient (ADC). This parameter provides additional clinical information about breast lesions, and potentially allows for in-vivo histological characterization. Purpose – To characterize and differentiate breast lesions through ADC analysis. Methodology – The study comprised 20 women, with 23 breast lesions: 3 benign lesions - 3 Fibroadenomas (FA); and 20 malignant - 16 Invasive Ductal Carcinomas (CDI), 2 Ductal Carcinomas In Situ (CDIS), 2 Invasive Lobular Carcinoma (CLI). On a 1.5T equipment a diffusion-weighted sequence with 6 b-values (b=0,50,250,500,750,1000 s/mm2) was used to examine the patients. ADC was obtained by fitting an exponential to data of lesion signal intensity vs. b values. A correspondence of ADC values to histological lesion characterization was done and finally, the results were comparison with the literature. Results and Discussion – Malignant lesions showed inferior ADCs significantly (p=0.014) lower ((0.94±0.22)x10-3 mm2/s) than the benign lesions ((1.43±0.25)x10-3 mm2/s). This may be associated to increasead cellularity in malignant lesions, which result in higher tissue restriction to diffusion. CDI showed low ADC values ((0.88±0.17)x10-3 mm2/s), while the CDIS showed higher ADCs ((1.33±0.29)x10-3 mm2/s). These results agree with the fact that CDIS are limited to mammary ducts, maintaining a less altered neighboring tissue structure, which results in a lower restriction to diffusion than observed in CDI. Significant differences between FA and CDI (p=0.010) and between CDI and CDIS (p=0.049) were observed. Conclusion – The ADC parameter is able to differentiate between malignant and benign breast lesions, as well as between some histological types.

Non-colorectal intestinal tract carcinomas in inflammatory bowel disease: results of the 3rd ECCO Pathogenesis Scientific Workshop (II)

Patients with inflammatory bowel diseases (IBD) have an excess risk of certain gastrointestinal cancers. Much work has focused on colon cancer in IBD patients, but comparatively less is known about other more rare cancers. The European Crohn's and Colitis Organization established a pathogenesis workshop to review what is known about these cancers and formulate proposals for future studies to address the most important knowledge gaps. This article reviews the current state of knowledge about small bowel adenocarcinoma, ileo-anal pouch and rectal cuff cancer, and anal/perianal fistula cancers in IBD patients.

TERT Promoter Mutations Are a Major Indicator of Poor Outcome in Differentiated Thyroid Carcinomas

Context: Telomerase promoter mutations (TERT) were recently described in follicular cell-derived thyroid carcinomas (FCDTC) and seem to be more prevalent in aggressive cancers. Objectives: We aimed to evaluate the frequency of TERT promoter mutations in thyroid lesions and to investigate the prognostic significance of such mutations in a large cohort of patients with differentiated thyroid carcinomas (DTCs). Design: This was a retrospective observational study. Setting and Patients: We studied 647 tumors and tumor-like lesions. A total of 469 patients with FCDTC treated and followed in five university hospitals were included. Mean follow-up (±SD) was 7.8 ± 5.8 years. Main Outcome Measures: Predictive value of TERT promoter mutations for distant metastasization, disease persistence at the end of follow-up, and disease-specific mortality. Results: TERT promoter mutations were found in 7.5% of papillary carcinomas (PTCs), 17.1% of follicular carcinomas, 29.0% of poorly differentiated carcinomas, and 33.3% of anaplastic thyroid carcinomas. Patients with TERT-mutated tumors were older (P < .001) and had larger tumors (P = .002). In DTCs, TERT promoter mutations were significantly associated with distant metastases (P < .001) and higher stage (P < .001). Patients with DTC harboring TERT promoter mutations were submitted to more radioiodine treatments (P = .009) with higher cumulative dose (P = .004) and to more treatment modalities (P = .001). At the end of follow-up, patients with TERT-mutated DTCs were more prone to have persistent disease (P = .001). TERT promoter mutations were significantly associated with disease-specific mortality [in the whole FCDTC (P < .001)] in DTCs (P < .001), PTCs (P = .001), and follicular carcinomas (P < .001). After adjusting for age at diagnosis and gender, the hazard ratio was 10.35 (95% confidence interval 2.01–53.24; P = .005) in DTC and 23.81 (95% confidence interval 1.36–415.76; P = .03) in PTCs. Conclusions: TERT promoter mutations are an indicator of clinically aggressive tumors, being correlated with worse outcome and disease-specific mortality in DTC. TERT promoter mutations have an independent prognostic value in DTC and, notably, in PTC.