The Met receptor tyrosine kinase (RTK) is expressed in the mammary gland under both normal and neoplastic conditions. Overexpression of the Met receptor is found in 15--20% of human breast cancers and is correlated with shortened disease-free interval and overall survival. In order to explore the role of dysregulated Met receptor signaling on the development of mammary tumors I have characterized a transgenic mouse model that expresses either wild type or a dysregulated Met receptor in the mammary epithelium under the control of the mouse mammary tumor virus promoter/enhancer (MMTV-Met). The Met receptor variants contained a mutation that results in decreased receptor ubiquitination and prolonged receptor signaling (Y1003F) or an activating mutation that was originally observed in patients with papillary renal carcinoma (M1250T) or both mutations (YF/MT). In vitro and in vivo transformation assays demonstrated that each mutation singly is weakly transforming, however, there was an additive effect on transformation when both mutations were present. This additive effect was observed in the transgenic mice where multiparous MMTV-Met-YF/MT mice developed tumors earlier and with much greater penetrance than did mice expressing either of the single mutants. This provides the first in vivo model that demonstrates a role for ubiquitination in suppression of transforming activity of an RTK. MMTV-Met-YF/MT tumors displayed a range of histological phenotypes but were mainly comprised of luminal lineage cells. Notably, MMTV-Met-M1250T tumors contained cells from both the basal and luminal populations, suggesting transformation of a progenitor cell. Progenitor cell transformation in RTK transgenic mouse models is uncommon and highlights distinct signaling differences and potentially lineage specificity of the two Met mutants. / Through assays of overexpression in vivo and inhibition in vitro, Met receptor signaling has been correlated with the development of the mammary gland. To examine the effects of loss of Met receptor signaling on mammary gland development I have utilized the Cre/LoxP1 recombination system to knock-out the Met receptor from the mammary epithelium. Mammary-specific Cre recombinase efficiently excised floxed DNA as visualized by activation of a beta-galactosidase reporter In Met+/+ glands, however, few beta-galactosidase positive cells are retained In the Mefl/fl glands and an intermediate number are retained in the Met fl/+ glands. This indicates that Met-null cells are selected against and supports a role for Met in the development of the mammary gland.

Thesis (Ph. D.)--Cornell University, January, 2009. / Vita. Includes bibliographical references.

Thesis (Ph. D.)--Cornell University, May, 2008. / Vita. Includes bibliographical references (leaves 115-128).

As modalidades terapêuticas para o câncer de mama têm promovido avanços no seu tratamento, levando ao aumento da sobrevida das mulheres. Contudo, com o aumento do tempo de vida, há também o risco do aumento de comorbidades relacionadas à idade, ao tratamento e à doença, sendo as principais: a obesidade, hipertensão arterial sistêmica (HAS) e diabetes mellitus (DM). Assim, realizou-se o presente estudo com o objetivo de identificar a prevalência de obesidade, HAS e níveis glicêmicos alterados entre mulheres com câncer de mama que frequentam um núcleo de reabilitação e avaliar a distribuição destas comorbidades em relação à idade, tempo de diagnóstico do câncer de mama, circunferência da cintura, hormonioterapia, tratamento quimioterápico e hábitos de vida. Trata-se de um estudo observacional, descritivo e transversal, no qual foram incluídas 67 mulheres com câncer de mama. Para a coleta de dados utilizou-se um instrumento com variáveis sociodemográficas e clínicas, além das avaliações antropométricas, pregas cutâneas, bioimpedância, pressão arterial sistêmica e coletado sangue para glicemia de jejum, no período de setembro a novembro de 2015. Foi realizada análise descritiva, resultando em tabelas de frequência para variáveis qualitativas; para identificar a relação entre as variáveis qualitativas ordinais ou nominais foi empregado o teste exato de Fischer e para estudar a relação entre as medidas contínuas e categóricas da pesquisa foram empregadas técnicas de estatística não paramétrica com o teste de Mann-Whitney (MW) e Kruskal-Wallis (KW). Em relação à obesidade, encontrou-se que 34,3% das participantes eram pré-obesas e 29,9% apresentavam índice de massa corpórea (IMC) com parâmetros de obesidade leve a grave; 53,7% eram hipertensas e 20,9% diabéticas. Encontrou-se associação entre idade e HAS; circunferência da cintura e HAS; níveis glicêmicos e circunferência da cintura; IMC e circunferência da cintura; idade e DM; e idade e circunferência da cintura. Os resultados encontrados mostram alta prevalência de obesidade, hipertensão arterial e níveis glicêmicos alterados nas mulheres participantes, assim como possibilitaram identificar os fatores associados às comorbidades, sendo estes a idade, hábitos de vida e tratamentos realizados para o câncer de mama / The therapeutic modalities for breast cancer have promoted advances in its treatment leading to increased survival of women. However, with the increase in longevity, there is also the risk of comorbidities related to age, to treatment and to the disease, the main ones being: obesity, systemic arterial hypertension (SAH) and diabetes mellitus (DM). Thus, the objective of the present study was to indentify the prevalence of obesity, SAH and altered glucose levels among women with breast câncer who attend a rehabilitation center and to evaluate the distribution of such comorbidities in relation to age, time of breast cancer diagnosis, waist circumference, hormone therapy, chemotherapy treatment and life habits. This is an observational, descriptive and transversal study, which included 67 women with breast cancer. An instrument with sociodemographic and clinical variables was used for the collection of data, in addition to anthropometric evaluations, skin folds, bioimpedance, systemic arterial pressure and blood collected for fasting blood glucose test, from September to November, 2015. A descriptive analysis was perfomed resulting in frequency tables for qualitative variables; the Fischer exact test was employed to identify the relationship between the ordinal or nominal qualitative variables, whereas non-parametric statistical techniques with the Mann-Whitney (MW) and Kruskal-Wallis (KW) test were employed to study the relationship between continuous and categorical measures of the survey. In relation to obesity, 34.3% of participants were found to be pre-obese, 29.9% had a body mass índex (BMI) with mild to severe obesity parameters, 53.7% were hypertensive and 20.9% had diabetes. Association was found between age and SAH; waist circumference and SAH; blood glucose levels and waist circunference; BMI and waist circunference; age and DM; and age and waist circunference. The findings show high prevalence of obesity, high blood pressure and altered glucose levels in the participating women, and allow for identification of the factors associated with comorbidities, namely age, life habits and breast cancer treatments

Submitted by (edna.saturno@ufrpe.br) on 2016-10-10T14:58:02Z No. of bitstreams: 1 Giuliana Viegas Schirato.pdf: 2698527 bytes, checksum: 33a822ba864e7a3747d0b9dd0d006691 (MD5) / Made available in DSpace on 2016-10-10T14:58:02Z (GMT). No. of bitstreams: 1 Giuliana Viegas Schirato.pdf: 2698527 bytes, checksum: 33a822ba864e7a3747d0b9dd0d006691 (MD5) Previous issue date: 2011-02-24 / After skin tumors and lymphohematopoietic tissue, mammary neoplasms are most common in domestic cats. Mammary carcinomas of cats have poor prognosis, since such tumors are very aggressive and with great ability to generate metastases. The aim of this study was to characterize the point of view histopathological, histochemical and immunohistochemical, spontaneously occurring mammary tumors in cats. Thirty-five samples of tumor tissue were collected surgically and histological sections were stained with hematoxylin-eosin (HE) for histopathological analysis. For histochemical analysis, were used the Masson’s Trichrome and Periodic Acid Schiff (PAS) to evaluate the deposition of collagen and glycosaminoglycans associated with tumors, respectively. To perform the immunohistochemistry assays were used anti-Ki-67, anti-PCNA and anti-Bcl-2. The obtained results showed that 14.29% of the lesions were classified as benign (fibroadenoma, adenoma and simple adenosis) and 85.71% as adenocarcinomas. Malignant neoplasms were multifocal, predominantly of invasive growth pattern, presence of necrosis, mitotic figures, and inflammatory infiltrate. The histochemical analysis revealed that the malignant lesions exhibited moderate to strong staining pattern when stained with PAS and weak to moderate with MT. Marking anti-Ki-67 showed cytoplasmic and nuclear pattern in well-differentiated malignant cells in tumor tissue. The protein Bcl-2 showed membrane pattern restricted to the processed tissue and the tissue marked with anti-PCNA showed nuclear and cytoplasmic pattern in glandular cells, occasionally in stromal cells of epithelioid appearance. In summary, the large number of mitotic figures and increased reactivity to staining with anti-PCNA antibody suggests neoplasms with intense proliferative activity indicating a poor prognosis for affected individuals. / Com exceção dos tumores de pele e do tecido linfo hematopoiético, o neoplasmas mamários são os mais comuns em felinos domésticos. Os carcinomas mamários de gatas possuem prognóstico desfavorável, uma vez que tais tumores são bastante agressivos e com grande capacidade de gerar metástases. Objetivou-se com esse estudo, caracterizar sob o ponto de vista histopatológico, histoquímico e imunoistoquímico; neoplasias mamárias de ocorrência espontânea em gatas. Trinta e cinco amostras de tecido neoplásico foram coletadas cirurgicamente e os cortes histológicos foram corados pela Hematoxilina-Eosina (HE), para estudo histopatológico. Para análise histoquímica, utilizou-se o Tricromico de Masson e Ácido Periódico de Schiff (PAS), para avaliar os depósitos de colágeno e glicosaminoglicanos associados aos tumores, respectivamente. Para realização dos ensaios imunoistoquímicos foram utilizados os anticorpos anti-Ki-67, anti-PCNA e anti-Bcl-2. Nos resultados obtidos observou-se que 14,29% das lesões foram classificadas como benignas (fibroadenoma, adenoma simples e adenose) e 85,71% como adenocarcinomas. As neoplasias malignas eram multifocais, de padrão de crescimento predominantemente invasivo, com presença de necrose, figuras de mitose, e infiltrado inflamatório. As análises histoquímicas revelaram que as lesões malignas apresentavam padrão de coloração moderada a forte quando coradas com PAS e fraca a moderada com Tricrômico de Masson. A marcação anti-Ki-67 apresentou padrão citoplasmático e nuclear em células malignas bem diferenciadas no tecido neoplásico. A proteína Bcl-2 apresentou padrão membranar restrito ao tecido transformado e o tecido marcado com o anticorpo anti-PCNA, demonstrou padrão citoplasmático e nuclear em células glandulares, ocasionalmente em células estromais de aspecto epitelióide. Em resumo, a grande quantidade de figuras de mitose e a maior reatividade à marcação com o anticorpo anti-PCNA sugere neoplasmas com intensa atividade proliferativa evidenciando um mau prognóstico para os indivíduos portadores.

No description available.

Os marcadores prognósticos em mastologia têm sido utilizados como apoio diagnóstico para prever o comportamento dos neoplasmas mamários (prognóstico) e determinar a provável resposta ao tratamento pré ou pós-cirúrgico. Estudos têm sido feitos sobre o prognóstico dos tumores mamários caninos (TMCs) que apresentam semelhanças e diferenças com tumores mamários humanos. Além disso, esses tumores exibem, com alta freqüência, proliferação de células mioepiteliais que podem sofrer metaplasia, acompanhada de alterações moleculares. O presente estudo teve o objetivo de verificar a expressão imuno-histoquímica e a associação de diferentes marcadores utilizados como prognósticos nos tumores de mama humana (RE, RP, c-erbB-2 e Ki-67) e marcadores mioepiteliais (p63, CK5 e vimentina) nos TMCs. O primeiro artigo analisa a expressão desses marcadores em 35 tumores encontrados em onze fêmeas caninas nas quais foram identificados tumores malignos múltiplos nas glândulas mamárias. Cada tipo histológico analisado em fêmeas portadoras de tumores múltiplos expressou marcadores prognósticos e mioepiteliais peculiares à sua histogênese, porém houve associação dessa expressão apenas em alguns tipos celulares presentes nos TMCs. Os tumores com componente epitelial carcinomatoso não apresentaram diferenças significativas, no entanto, nos tumores com componente complexo e misto, ocorreu associação entre a expressão da p63, CK5 e vimentina. No conjunto de marcadores estudados, a p63 e a CK5 mostram-se promissoras na elucidação da transformação das células mioepiteliais concomitante à invasão tumoral e com relação à expressão da vimentina que se mostrou bem evidenciada durante a transformação da célula mioepitelial proliferada a célula participante do mesênquima do neoplasma invasor em mamas de caninos, ao menos nos aspectos de expressão molecular e morfológica. O segundo artigo analisa os marcadores mioepiteliais em 82 casos de TMCs malignos. Este estudo comprovou a freqüência e a associação da expressão desses marcadores em determinados tipos histológicos tumorais e celulares, permitindo a identificação das células mioepiteliais em transformação na maior parte dos TMCs malignos, notadamente, os que apresentam componente mesenquimal metaplásico. Mais estudos devem ser feitos na tentativa de verificar a significância da expressão encontrada no comportamento biológico desses tumores. / Prognostic markers in mastology have been used as diagnostic support, to predict the behavior of mammary neoplasias (prognosis) and to determine their possible response to treatment before or after surgery. Studies have been conducted on the prognosis of canine mammary gland tumors (MGTs), which show similarities to and differences from human breast tumors. In addition, these tumors often show proliferation of myoepithelial cells, which may undergo metaplasia, accompanied by molecular alterations. The aim of the present study was to check the immunohistochemical expression and the association between different markers used as prognostic factors in human breast tumors (ER, RP, c-erbB-2 and Ki-67) and myoepithelial markers (p63, CK5 and vimentin) in MGTs. The first article analyzes the expression of these markers in 35 tumors in 11 female dogs, where multiple tumors were identified in the mammary glands. Each histological type analyzed in the female dogs with multiple tumors expressed prognostic and myoepithelial markers that were peculiar to their histogenesis, but the association of this expression was observed only in some cell types of MGTs. Tumors with a carcinomatous epithelial component did not have significant differences, but tumors with complex and mixed components showed association between the expressions of p63, CK5 and vimentin. Of the group of investigated markers, p63 and CK5 proved to be promising tools in elucidating the transformation of myoepithelial cells concomitantly to tumor invasion and in terms of vimentin expression, which was quite pronounced in this transformation from proliferating myoepithelial cells into cells that participate in the mesenchyma of the invasive neoplasia in canine mammary glands, at least with regard to the aspects of molecular and morphological expression. The second article analyzes myoepithelial markers in 82 cases of malignant MGTs. This study corroborated the frequency and association of the expression of these markers in certain histological tumor and cell types, allowing for the identification of myoepithelial cells in transformation in most malignant MGTs, chiefly those with a metaplastic mesenchymal component. Further studies are necessary in order to assess the importance of expression found in the biological behavior of these tumors.

A interação entre as vias de sinalização dos estrógenos e fatores de crescimento está relacionada a maior agressividade dos tumores de mama. Assim, o objetivo deste trabalho foi verificar a interação entre as vias do E2 e do EGF no controle da expressão dos genes HNRPK, PAWR e PHLDA1 nas células MCF-7 (ER+) e MDA-MB-231 (ER-). Nas células MCF-7, o EGF e o E2 diminuíram a expressão do PAWR e aumentaram a expressão de PHLDA1. A inibição do ER pelo ICI resultou no aumento da expressão de PAWR sendo que a adição do E2 ou do EGF diminuiu sua expressão sem retomar os níveis observados nos tratamentos com E2 ou EGF isoladamente. Para o gene PHLDA1, o efeito do E2 e do EGF não variou após o tratamento com ICI. Nas células MDA-MB-231, a ação do EGF foi mais efetiva sobre a expressão de PAWR. A via ERK1/2 é importante na ativação do ER pelo EGF. O efeito do EGF sobre o PHLDA1 ocorre através da ativação das vias ERK1/2 e p38 MAPK. Estes resultados mostram a interação entre as vias do E2 e do EGF no controle da expressão do PAWR, mas não do PHLDA1. / The crosstalk between estrogens and growth factors pathways has been associated with breast cancer aggressiveness. Based on this, the present study aimed to determine the possible crosstalk between E2 and EGF pathways on the HNRPK, PAWR and PHLDA1 expression regulation in MCF-7 (ER+) and MDA-MB-231 (ER-) cells. In MCF-7 cells, treatments with E2 and EGF decreased PAWR expression and increased PHLDA1 expression. The ER inhibition by the ICI treatment resulted in increased PAWR expression. The E2 or EGF addition down-regulated its expression without a return to the levels observed after the E2 or EGF treatments alone. To the PHLDA1 gene, the effect of E2 and EGF treatments did not change after the ICI treatment. In MDA-MB-231 cells, the EGF effect was more significant on the PAWR gene expression control. The ERK1/2 pathway is important to the ER activation by EGF. The EGF effect over the PHLDA1 is dependent on the ERK1/2 and p38 MAPK activation. These findings suggest a crosstalk between E2 and EGF pathways on the PAWR expression control but not to PHLDA1.

Vitamin D3 must be metabolically activated by the liver to 25-hydroxyvitamin D3 (25OHD3) and then by the kidney 1alphahydroxylase (1alphaOHase) to become 1,25dihydroxyvitamin D 3 (1,25(OH)2D3). 1,25(OH)2 D3 is a potent inhibitor of tumor growth in vitro and in vivo. Recent studies indicate that metabolic activation of 1,25(OH) 2D3 also occurs in cancer cells such as breast cancer. Consequently, the major objective of this project was to determine if tumoral 25OHD 3-1alphahydroxylase modulates any or all of the stages of breast tumor progression without inducing the hypercalcemic side effects of 1,25(OH) 2D3. For this purpose we used the PyVMT breast cancer mouse model in which the oncoprotein, polyomamiddle T antigen (PyMT) is under the control of mouse mammary tumor virus LTR (MMTV LTR). Mice exhibited tumors restricted to the mammary epithelium progressing to the various stages of breast cancer. Animals were treated with either vehicle, 25OHD3 (2000 pM/24h) or 1,25(OH)2D3 (12pM/24h). Mice treated with the vitamin D precursor, 25OHD3, exhibited a marked reduction in tumor onset and growth comparable to the 1,25(OH)2D3 treated group. Furthermore, biomarkers of tumor progression were markedly reduced in 25OHD3 and 1,25(OH)2D3 animals as compared to vehicle-treated animals. However, mean circulating calcium concentrations remained unchanged in 25OHD3 treated animals but increased significantly in 1,25(OH)2D3 treated animals as compared to controls. Tumoral levels of 1,25(OH)2D3 in mice treated with 25OHD3 were increased 79% in comparison to vehicle control mice. Additionally, 25OHD3 and 1,25(OH)2D 3 treated animals had a significant decrease in the mean number of lung metastases per animal as compared to vehicle treated control animals. This study therefore suggests an important autocrine role of 1alphaOHase expression in breast tumor cells. Furthermore, accumulation of intra-tumoral 1,25(OH) 2D3 in response to 25OHD3 administration strongly suggests that locally produced 1,25(OH)2D3 plays a significant role in restraining tumor growth without inducing the hypercalcemic side effects associated with 1,25(OH)2D3.

Os marcadores prognósticos em mastologia têm sido utilizados como apoio diagnóstico para prever o comportamento dos neoplasmas mamários (prognóstico) e determinar a provável resposta ao tratamento pré ou pós-cirúrgico. Estudos têm sido feitos sobre o prognóstico dos tumores mamários caninos (TMCs) que apresentam semelhanças e diferenças com tumores mamários humanos. Além disso, esses tumores exibem, com alta freqüência, proliferação de células mioepiteliais que podem sofrer metaplasia, acompanhada de alterações moleculares. O presente estudo teve o objetivo de verificar a expressão imuno-histoquímica e a associação de diferentes marcadores utilizados como prognósticos nos tumores de mama humana (RE, RP, c-erbB-2 e Ki-67) e marcadores mioepiteliais (p63, CK5 e vimentina) nos TMCs. O primeiro artigo analisa a expressão desses marcadores em 35 tumores encontrados em onze fêmeas caninas nas quais foram identificados tumores malignos múltiplos nas glândulas mamárias. Cada tipo histológico analisado em fêmeas portadoras de tumores múltiplos expressou marcadores prognósticos e mioepiteliais peculiares à sua histogênese, porém houve associação dessa expressão apenas em alguns tipos celulares presentes nos TMCs. Os tumores com componente epitelial carcinomatoso não apresentaram diferenças significativas, no entanto, nos tumores com componente complexo e misto, ocorreu associação entre a expressão da p63, CK5 e vimentina. No conjunto de marcadores estudados, a p63 e a CK5 mostram-se promissoras na elucidação da transformação das células mioepiteliais concomitante à invasão tumoral e com relação à expressão da vimentina que se mostrou bem evidenciada durante a transformação da célula mioepitelial proliferada a célula participante do mesênquima do neoplasma invasor em mamas de caninos, ao menos nos aspectos de expressão molecular e morfológica. O segundo artigo analisa os marcadores mioepiteliais em 82 casos de TMCs malignos. Este estudo comprovou a freqüência e a associação da expressão desses marcadores em determinados tipos histológicos tumorais e celulares, permitindo a identificação das células mioepiteliais em transformação na maior parte dos TMCs malignos, notadamente, os que apresentam componente mesenquimal metaplásico. Mais estudos devem ser feitos na tentativa de verificar a significância da expressão encontrada no comportamento biológico desses tumores. / Prognostic markers in mastology have been used as diagnostic support, to predict the behavior of mammary neoplasias (prognosis) and to determine their possible response to treatment before or after surgery. Studies have been conducted on the prognosis of canine mammary gland tumors (MGTs), which show similarities to and differences from human breast tumors. In addition, these tumors often show proliferation of myoepithelial cells, which may undergo metaplasia, accompanied by molecular alterations. The aim of the present study was to check the immunohistochemical expression and the association between different markers used as prognostic factors in human breast tumors (ER, RP, c-erbB-2 and Ki-67) and myoepithelial markers (p63, CK5 and vimentin) in MGTs. The first article analyzes the expression of these markers in 35 tumors in 11 female dogs, where multiple tumors were identified in the mammary glands. Each histological type analyzed in the female dogs with multiple tumors expressed prognostic and myoepithelial markers that were peculiar to their histogenesis, but the association of this expression was observed only in some cell types of MGTs. Tumors with a carcinomatous epithelial component did not have significant differences, but tumors with complex and mixed components showed association between the expressions of p63, CK5 and vimentin. Of the group of investigated markers, p63 and CK5 proved to be promising tools in elucidating the transformation of myoepithelial cells concomitantly to tumor invasion and in terms of vimentin expression, which was quite pronounced in this transformation from proliferating myoepithelial cells into cells that participate in the mesenchyma of the invasive neoplasia in canine mammary glands, at least with regard to the aspects of molecular and morphological expression. The second article analyzes myoepithelial markers in 82 cases of malignant MGTs. This study corroborated the frequency and association of the expression of these markers in certain histological tumor and cell types, allowing for the identification of myoepithelial cells in transformation in most malignant MGTs, chiefly those with a metaplastic mesenchymal component. Further studies are necessary in order to assess the importance of expression found in the biological behavior of these tumors.