Resumo: Com o objetivo de caracterizar pelo método histológico e imunoistoquimíco o câncer de mama de cadelas sem raça definida (SRD) foram estudados 40 animais com distribuição uniforme nos estágios I, II, III e IV da doença. Todos os animais foram submetidos a anamnese, exame físico e complementares (Citopatológico, Raio x, Ultra Som) para o estadiamento clínico do câncer (TNMc), amostra histológica da lesão para caracterização do tipo de tumor e amostra imunoistoquímica para determinação molecular das células em apoptose (caspase 3) e proliferação (PCNA). A amostra foi caracterizada com predominância de idade acima de seis anos (92,5%), obesos (57,5%) com ingestão de dieta mista (80%) caseira-industrial. 80% das cadelas já tiveram cria, apresentando pseudogestação (40%), tumoração das mamas inguinais (70%), em ambas as cadeias, com nódulos multicêntricos (82,5%) de incidência bilateral (60%), prevalecendo nódulos de tamanho inferior a 2cm de diâmetro (37,5%). Prevaleceu o carcinoma simples (70%) em relação ao carcinoma complexo (30%). Os animais com carcinoma simples apresentaram características de porte grande (50%), obesidade (57%), cios regulares (89%) e multíparas (82%) e prevalência idêntica entre os estadiamentos clínico (70%). O estadio mais grave (IV) esteve associado aos animais com idade média de 8 anos, ausência de obesidade (60%). A imunomarcação de PCNA e caspase-3 não apresentou diferença significativa entre os tipos de tumor, suas características clínicas e grau histológico, no entanto a caspase-3 teve associação significativa entre o estadiamento clínico (I<III e II<III) e presença de ulceração tumoral (P<0,049) / Abstract: The severity of breast cancer in mongrel animals is not defined yet in its clinical, histologic and imunoistoquimical associative aspects. With the aim to establish that characterization we studied 40 bitches, 3-15 years old, mixed breed attended at the U. Vet. Hospital ambulatories in compliance with current ethical and pathological rules. The animals were alloted according to their clinical stage in groups stage I, II, III and IV with 10 animals each. After physical examination and lifestyle records. The tumor nodules were evaluated macro-and-microscopically, the later through histopathologicaly and imunoistopathologicaly. From these data the animals were classified as predominantly up to six years old (92,5%), obese (57,5%), presenting simple carcinoma (CS) more frequent (70%) than complex carcinoma type (30%). 80% of the animals wasanþt nuliparus, showing pseudopregnancies (40%), having multicentric nodules (82,5%) sizing less than two cm diameter (37,5%), located inguinally (70%) in both side mammary chains. The immunohistochemistry label for PCNA and caspase-3 were not associated with tumors type, clinical aspects and histological grade, however only caspase-3 had significant association with clinical stage (I<III and II<III) and tumors ulceration (P<0,049) / Orientador: Enio Pedone Bandarra / Coorientador: Renée Laufer Amorim / Banca: Maria Denise Lopes / Banca: Luiz Henrique do Araujo Machado / Banca: Paulo César Maiorka / Banca: Andrigo Barbosa de Nardi / Doutor

Made available in DSpace on 2014-06-11T19:31:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-07-10Bitstream added on 2014-06-13T19:01:36Z : No. of bitstreams: 1 burini_chp_dr_botfmvz.pdf: 788656 bytes, checksum: 29e369ee0de982c1e6a5dbda2bc2c81c (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Com o objetivo de caracterizar pelo método histológico e imunoistoquimíco o câncer de mama de cadelas sem raça definida (SRD) foram estudados 40 animais com distribuição uniforme nos estágios I, II, III e IV da doença. Todos os animais foram submetidos a anamnese, exame físico e complementares (Citopatológico, Raio x, Ultra Som) para o estadiamento clínico do câncer (TNMc), amostra histológica da lesão para caracterização do tipo de tumor e amostra imunoistoquímica para determinação molecular das células em apoptose (caspase 3) e proliferação (PCNA). A amostra foi caracterizada com predominância de idade acima de seis anos (92,5%), obesos (57,5%) com ingestão de dieta mista (80%) caseira-industrial. 80% das cadelas já tiveram cria, apresentando pseudogestação (40%), tumoração das mamas inguinais (70%), em ambas as cadeias, com nódulos multicêntricos (82,5%) de incidência bilateral (60%), prevalecendo nódulos de tamanho inferior a 2cm de diâmetro (37,5%). Prevaleceu o carcinoma simples (70%) em relação ao carcinoma complexo (30%). Os animais com carcinoma simples apresentaram características de porte grande (50%), obesidade (57%), cios regulares (89%) e multíparas (82%) e prevalência idêntica entre os estadiamentos clínico (70%). O estadio mais grave (IV) esteve associado aos animais com idade média de 8 anos, ausência de obesidade (60%). A imunomarcação de PCNA e caspase-3 não apresentou diferença significativa entre os tipos de tumor, suas características clínicas e grau histológico, no entanto a caspase-3 teve associação significativa entre o estadiamento clínico (I<III e II<III) e presença de ulceração tumoral (P<0,049) / The severity of breast cancer in mongrel animals is not defined yet in its clinical, histologic and imunoistoquimical associative aspects. With the aim to establish that characterization we studied 40 bitches, 3-15 years old, mixed breed attended at the U. Vet. Hospital ambulatories in compliance with current ethical and pathological rules. The animals were alloted according to their clinical stage in groups stage I, II, III and IV with 10 animals each. After physical examination and lifestyle records. The tumor nodules were evaluated macro-and-microscopically, the later through histopathologicaly and imunoistopathologicaly. From these data the animals were classified as predominantly up to six years old (92,5%), obese (57,5%), presenting simple carcinoma (CS) more frequent (70%) than complex carcinoma type (30%). 80% of the animals wasanþt nuliparus, showing pseudopregnancies (40%), having multicentric nodules (82,5%) sizing less than two cm diameter (37,5%), located inguinally (70%) in both side mammary chains. The immunohistochemistry label for PCNA and caspase-3 were not associated with tumors type, clinical aspects and histological grade, however only caspase-3 had significant association with clinical stage (I<III and II<III) and tumors ulceration (P<0,049)

Orientador: Debora Aparecida Pires de Campos Zuccari / Banca: Mirela Tinucci Costa / Banca: Claudia Regina Bonini Domingos / Resumo: O estudo dos tumores mamários em cadelas revela-se um excelente modelo para a investigação clínico-patológica, diagnóstica e prognóstica de neoplasias mamárias. As interleucinas desempenham um papel fundamental no câncer, em especial a interleucina-8 (IL-8), que possui propriedades tumorigênicas e pró-angiogênicas e a interleucina-12 (IL-12), com propriedades anti-metastáticas e anti-angiogênicas. O marcador tumoral antígeno-carboidrato (CA 15-3) tem importante significado clínico no seguimento de pacientes com neoplasia mamária. Além disso, o DNA livre circulante de células tem sido considerado um candidato a biomarcador para tumores. Assim, foram objetivos deste estudo mensurar os níveis séricos das proteínas IL-8, IL-12, CA 15-3, bem como estimar o número de cópias das sequências CAN SINEs relacionando-os aos parâmetros clínico-patológicos e a sobrevida do grupo a fim de inferir o possível valor prognóstico desses marcadores para o câncer de mama. Através do ensaio imunoenzimático e da técnica de PCR em tempo real foram avaliadas 33 cadelas com neoplasia mamária e 50 cadelas controle e suas concentrações foram estatisticamente relacionadas aos parâmetros estabelecidos. Foi encontrada diferença significante entre os níveis séricos de IL-8 nas cadelas com neoplasia mamária quando comparadas ao grupo controle e além disso, verificou-se aumento de IL-8 em relação à evolução tumoral, ao envolvimento linfonodal, recidiva e óbito. Baixos níveis de IL-12 foram observados em cadelas com tumores com maior tempo de evolução, cadelas com idade superior a 10 anos e, além disso, a alta taxa de sobrevida teve correlação com níveis elevados de IL-12. Assim, o aumento da IL-8 e a diminuição da IL-12 podem ser considerados marcadores prognósticos independentes confirmando seu papel como citocina... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract:The study of mammary tumors in dogs appears to be an excellent model to investigate the clinical and pathological diagnosis and prognosis of breast cancer. Interleukins play a key role in cancer, particularly interleukin-8 (IL-8), which have tumorigenic and pro-angiogenic properties and interleukin-12 (IL-12), with anti-metastatic and anti-angiogenic properties. The tumor marker carbohydrate antigen (CA 15-3) also has important clinical significance in monitoring patients with breast cancer. In addition, free circulating DNA has been considered a candidate biomarker for cancer. The objectives of this study were to measure serum levels of protein IL-8, IL-12, CA 15-3 and to estimate the number of copies of CAN SINEs sequences relating them with clinicopathological parameters and survival of the group, in order to infer a likely value as prognostic markers in breast cancer. Through enzyme immunoassay and real-time PCR were evaluated 33 dogs with mammary cancer and 50 control dogs and their concentrations were statistically related with these parameters. Significant difference was found between IL-8 serum levels in bitches with mammary neoplasia in relation to the control group and in relation to disease progression, the lymph node involvement and in relation to death. Low levels of IL-12 were observed in female dogs with tumors with a longer evolution, aged 10 years and in addition, the high survival rate was correlated with high levels of IL-12. Thus, increased IL- 8 and decreased IL-12 can be considered independent prognostic markers which confirm their role as tumorigenic and anti-tumorigenic cytokines, respectively. For the CA 15-3 were found higher levels in dogs with lymph node involvement and poor survival of the group. In estimating the number of copies of CANSINEs, significant difference was found regarding the parameters indicators of poor prognosis and low survival. Our results... (Complete abstract click electronic access below) / Mestre

A neoplasia mamária é a doença mais entre as neoplasias em cadelas. As características clínicas dos tumores mamários caninos e sua relação com prognóstico foram discutidos, incluindo idade, raça, estagiamento clínico, diagnóstico histopatológico, hormônios e proliferação celular. Fatores clínicos prognósticos incluindo diâmetro do tumor e comprometimento linfonodal são discutidos em relação a graduação histopatológica e expressão de OCT4 (marcador para célula tronco tumoral). Avaliação imunohistoquímica dos marcadores (RE, RP, Ki-67 e OCT4) de neoplasias mamárias foi descritas. Foi observada marcação positiva para o OCT4, porém não se observou associação com fatores prognósticos clínicos. Assim como a marcação de RE, PR e Ki-67 foram observados. Houve uma forte associação entre graduação histopatológica de malignidade e tipo histológico. / Mammary neoplasms are the most common neoplasm in female dogs. The clinical features of canine mammary gland tumors and their relation to prognosis were discussed, including age, breed, staging, histopathological diagnosis, hormones, cell proliferation. Additional clinical prognostic factors including tumor size, and lymph node status are discussed in relation to graduation histopathological and OCT4 expression (cancer stem cell marker). Immunohistochemical evaluation of the markers (ER, PR, KI-67, OCT4) of the neoplastic canine mammary gland is described. We observed positive staining for OCT4, but was not associated with clinical prognostic factors. And the marking of ER, PR and Ki-67 was observed. There was a strong association between histopathological grade and histological type of malignancy.

Em animais, a prevalência do câncer tem aumentado de forma significativa com o passar dos anos. As neoplasias mamárias representam o tipo mais frequente de câncer em cadelas, chegando a 52% da população de fêmeas, e entre os animais afetados, 50% das neoplasias se apresentam sob a forma maligna. O desenvolvimento e caracterização de modelos animais para o estudo de neoplasias humanas é de extrema relevância para a melhoria no diagnóstico e tratamento do câncer. Os tumores sólidos apresentam uma hierarquia entre as células que determina o desenvolvimento e o comportamento da neoplasia. Recentemente, tem-se estudado um pequeno grupo de células que apresentam diversas características das células-tronco normais encontradas nos tecidos. Estas células, denominadas Células Iniciadoras de Tumor (CITs), são descritas como sendo as principais responsáveis pelas falhas na quimioterapia e no aparecimento de recidivas tumorais, devido ao grande potencial de renovação e diferenciação que elas possuem. Desta maneira, nosso objetivo foi caracterizar linhagens celulares provenientes de neoplasia de glândula mamária de cadelas que pudessem ser utilizadas futuramente na pesquisa básica e aplicada em oncologia comparada. Além das alterações citogenéticas e imunocitoquímicas, verificamos que os cultivos celulares apresentaram quantidades distintas de populações positivas e negativas quanto à expressão de CD24 e CD44 bem como da atividade de aldeído-desidrogenase, porém, todos exibiram potencial tumorigênico in vitro através do ensaio de formação de tumoresferas / The prevalence of cancer in animals has increased significantly over the years. Mammary tumors are the most common cancer in dogs, reaching 52% of female population, and among affected animals, 50% of tumors are presented in the malignant form. The development and characterization of animal models for the study of human cancers is extremely important for improving the diagnosis and treatment of cancer. Solid tumors have a hierarchy of cells that determines the development and behavior of cancer. Recently, there has been studied a small group of cells with different characteristics from those normal stem cells found in tissues. These cells, known as tumor initiating cells (TICs), are described as being primarily responsible for the failures in chemotherapy and the appearance of recurrences, because of their potential for renewal and differentiation. Thus, our objective was to characterize cell lines derived from mammary gland neoplasia of dogs that could be further used for basic and applied research in comparative oncology. In addition to the cytogenetic and immunocytochemical changes, we found that cell cultures had different amounts of positive and negative populations of CD24 and CD44 expression and aldehyde dehydrogenase activity, however, all exhibited tumorigenic potential in vitro through tumorspheres formation

Em animais, a prevalência do câncer tem aumentado de forma significativa com o passar dos anos. As neoplasias mamárias representam o tipo mais frequente de câncer em cadelas, chegando a 52% da população de fêmeas, e entre os animais afetados, 50% das neoplasias se apresentam sob a forma maligna. O desenvolvimento e caracterização de modelos animais para o estudo de neoplasias humanas é de extrema relevância para a melhoria no diagnóstico e tratamento do câncer. Os tumores sólidos apresentam uma hierarquia entre as células que determina o desenvolvimento e o comportamento da neoplasia. Recentemente, tem-se estudado um pequeno grupo de células que apresentam diversas características das células-tronco normais encontradas nos tecidos. Estas células, denominadas Células Iniciadoras de Tumor (CITs), são descritas como sendo as principais responsáveis pelas falhas na quimioterapia e no aparecimento de recidivas tumorais, devido ao grande potencial de renovação e diferenciação que elas possuem. Desta maneira, nosso objetivo foi caracterizar linhagens celulares provenientes de neoplasia de glândula mamária de cadelas que pudessem ser utilizadas futuramente na pesquisa básica e aplicada em oncologia comparada. Além das alterações citogenéticas e imunocitoquímicas, verificamos que os cultivos celulares apresentaram quantidades distintas de populações positivas e negativas quanto à expressão de CD24 e CD44 bem como da atividade de aldeído-desidrogenase, porém, todos exibiram potencial tumorigênico in vitro através do ensaio de formação de tumoresferas / The prevalence of cancer in animals has increased significantly over the years. Mammary tumors are the most common cancer in dogs, reaching 52% of female population, and among affected animals, 50% of tumors are presented in the malignant form. The development and characterization of animal models for the study of human cancers is extremely important for improving the diagnosis and treatment of cancer. Solid tumors have a hierarchy of cells that determines the development and behavior of cancer. Recently, there has been studied a small group of cells with different characteristics from those normal stem cells found in tissues. These cells, known as tumor initiating cells (TICs), are described as being primarily responsible for the failures in chemotherapy and the appearance of recurrences, because of their potential for renewal and differentiation. Thus, our objective was to characterize cell lines derived from mammary gland neoplasia of dogs that could be further used for basic and applied research in comparative oncology. In addition to the cytogenetic and immunocytochemical changes, we found that cell cultures had different amounts of positive and negative populations of CD24 and CD44 expression and aldehyde dehydrogenase activity, however, all exhibited tumorigenic potential in vitro through tumorspheres formation

A neoplasia mamária é a doença mais entre as neoplasias em cadelas. As características clínicas dos tumores mamários caninos e sua relação com prognóstico foram discutidos, incluindo idade, raça, estagiamento clínico, diagnóstico histopatológico, hormônios e proliferação celular. Fatores clínicos prognósticos incluindo diâmetro do tumor e comprometimento linfonodal são discutidos em relação a graduação histopatológica e expressão de OCT4 (marcador para célula tronco tumoral). Avaliação imunohistoquímica dos marcadores (RE, RP, Ki-67 e OCT4) de neoplasias mamárias foi descritas. Foi observada marcação positiva para o OCT4, porém não se observou associação com fatores prognósticos clínicos. Assim como a marcação de RE, PR e Ki-67 foram observados. Houve uma forte associação entre graduação histopatológica de malignidade e tipo histológico. / Mammary neoplasms are the most common neoplasm in female dogs. The clinical features of canine mammary gland tumors and their relation to prognosis were discussed, including age, breed, staging, histopathological diagnosis, hormones, cell proliferation. Additional clinical prognostic factors including tumor size, and lymph node status are discussed in relation to graduation histopathological and OCT4 expression (cancer stem cell marker). Immunohistochemical evaluation of the markers (ER, PR, KI-67, OCT4) of the neoplastic canine mammary gland is described. We observed positive staining for OCT4, but was not associated with clinical prognostic factors. And the marking of ER, PR and Ki-67 was observed. There was a strong association between histopathological grade and histological type of malignancy.

Abstract Peroxisome proliferator-activated receptors (PPARs) are transcription factors that belong to the family of nuclear hormone receptors and exist as three isoforms namely PPARα, PPARβ and PPARγ. PPARs function as key regulators of glucose and lipid metabolism and are potential targets for drugs used in the treatment of glucose and lipid metabolism dysregulation. PPARs also regulate the expression of genes involved in the process of cellular proliferation and differentiation. Since it was discovered that PPAR ligands cause liver tumourigenesis in rodents, PPARs and their modulators have been investigated widely in in vitro and in vivo studies of carcinogenesis of the liver, colon, prostate, lung and skin. PPARα and PPARγ are the most studied PPAR isoforms in relation to cancer, while the association of PPARβ with cancer is increasingly being investigated. Some studies suggest that PPARβ and its ligands may have anticancer activity, while other studies identify a role for PPARβ in tumour promotion and progression. Breast cancer is one of the most common types of cancer in women with the majority caused by non-hereditary mechanisms. The activation of PPARα in breast cancer cells is associated with an increase in proliferation, while PPARγ activation in breast cancer cells is related to differentiation and an inhibition of cell proliferation. The role of PPARβ and its modulators in breast cancer is uncertain, as there have been limited studies addressing the effects of PPARβ modulation in breast cancer cell lines. Environmental contaminants such as the phthalate plasticizers and alcohol are putative risk factors for breast cancer. The phthalates di(2-ethylhexyl)phthalate (DEHP) and di-n-butyl phthalate (DBP) are plasticizers that are used in a range of common household, medical and beauty products and as a consequence humans are exposed to significant levels of these compounds. DEHP and DBP are known teratogens in rodents and DEHP induces hepatocarcinogenesis in a process thought to be mediated via PPARα. DEHP and DBP are metabolized in vivo by esterases to the monoesters, mono-(2-ethylhexyl) phthalate (MEHP) and mono-n-butyl phthalate (MBP), and these compounds have been identified in human biological samples. MEHP and MBP modulate PPARs in various tissues and cell types, but their ability to modulate PPARs in human breast cancer cells is not known. Like phthalates, ethanol is another modulator of PPARs and alcohol consumption is associated positively with breast cancer development, but the molecular mechanisms involved are unknown and there are no studies that examine the effects of ethanol and its metabolite acetaldehyde on PPARs in breast cancer cell lines. This thesis describes studies establishing and validating a breast cancer cell line that conditionally expresses human PPARβ under the control of a tetracycline regulator. Using this model, the ability of PPARβ over-expression and/or activation by the PPARβ specific ligand GW0742 to promote breast cancer cell proliferation was studied. Furthermore, putative PPARβ regulated genes were examined for alterations in expression in the presence of the PPARβ ligand. This work determined that over-expression of PPARβ and/or its activation by GW0742 does not promote proliferation in MCF-7 breast cancer cells. This thesis also investigated the effects of the phthalate monoesters MEHP and MBP on PPARs in MCF-7 breast cancer cells. It was found that MEHP activated both PPARα and PPARγ but was unable to activate PPARβ, whereas MBP could not activate any of the PPAR isoforms. MBP was an antagonist for both PPARγ and PPARβ. Using breast cancer cell lines, studies were conducted addressing the effects of an increasing concentration of ethanol (0-300 mM) on the transcription and transactivation of PPARα and PPARβ isoforms. Estrogen receptor positive MCF-7 breast cancer cells were more sensitive to the effects of ethanol than estrogen receptor negative MDA-MB-231 cells, with changes in PPARα mRNA more pronounced than PPARβ mRNA. Studies in MCF-7 cells conditionally expressing either PPARα or PPARβ in the presence of their respective specific ligands, GW7647 and GW0742, revealed that ethanol concentrations of 20 mM and 100 mM suppressed the maximal response to ligand-mediated activation for PPARα. Studies using the ethanol metabolism enzyme inhibitors 4-methylpyrazole and cyanamide, suggested that while ethanol was responsible for the modulation of PPARβ transactivation, the primary metabolite acetaldehyde was responsible for the effects on PPARα transactivation. Lastly, it was determined that ethanol and/or GW0742 did not increase the proliferation of MCF-7 Tet-off cells. The findings in this thesis suggest that given the different consequences of MEHP, MBP and ethanol on PPARs, PPAR expression and activation by ligands may have tissue specific consequences and that PPARβ may have a complex role in mammary gland tumourigenesis.

Made available in DSpace on 2014-06-11T19:26:05Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-12Bitstream added on 2014-06-13T18:53:59Z : No. of bitstreams: 1 gelaleti_gb_me_sjrp.pdf: 706621 bytes, checksum: f280ddfe72d9aced3754ba3a7a380f9c (MD5) / O estudo dos tumores mamários em cadelas revela-se um excelente modelo para a investigação clínico-patológica, diagnóstica e prognóstica de neoplasias mamárias. As interleucinas desempenham um papel fundamental no câncer, em especial a interleucina-8 (IL-8), que possui propriedades tumorigênicas e pró-angiogênicas e a interleucina-12 (IL-12), com propriedades anti-metastáticas e anti-angiogênicas. O marcador tumoral antígeno-carboidrato (CA 15-3) tem importante significado clínico no seguimento de pacientes com neoplasia mamária. Além disso, o DNA livre circulante de células tem sido considerado um candidato a biomarcador para tumores. Assim, foram objetivos deste estudo mensurar os níveis séricos das proteínas IL-8, IL-12, CA 15-3, bem como estimar o número de cópias das sequências CAN SINEs relacionando-os aos parâmetros clínico-patológicos e a sobrevida do grupo a fim de inferir o possível valor prognóstico desses marcadores para o câncer de mama. Através do ensaio imunoenzimático e da técnica de PCR em tempo real foram avaliadas 33 cadelas com neoplasia mamária e 50 cadelas controle e suas concentrações foram estatisticamente relacionadas aos parâmetros estabelecidos. Foi encontrada diferença significante entre os níveis séricos de IL-8 nas cadelas com neoplasia mamária quando comparadas ao grupo controle e além disso, verificou-se aumento de IL-8 em relação à evolução tumoral, ao envolvimento linfonodal, recidiva e óbito. Baixos níveis de IL-12 foram observados em cadelas com tumores com maior tempo de evolução, cadelas com idade superior a 10 anos e, além disso, a alta taxa de sobrevida teve correlação com níveis elevados de IL-12. Assim, o aumento da IL-8 e a diminuição da IL-12 podem ser considerados marcadores prognósticos independentes confirmando seu papel como citocina... / The study of mammary tumors in dogs appears to be an excellent model to investigate the clinical and pathological diagnosis and prognosis of breast cancer. Interleukins play a key role in cancer, particularly interleukin-8 (IL-8), which have tumorigenic and pro-angiogenic properties and interleukin-12 (IL-12), with anti-metastatic and anti-angiogenic properties. The tumor marker carbohydrate antigen (CA 15-3) also has important clinical significance in monitoring patients with breast cancer. In addition, free circulating DNA has been considered a candidate biomarker for cancer. The objectives of this study were to measure serum levels of protein IL-8, IL-12, CA 15-3 and to estimate the number of copies of CAN SINEs sequences relating them with clinicopathological parameters and survival of the group, in order to infer a likely value as prognostic markers in breast cancer. Through enzyme immunoassay and real-time PCR were evaluated 33 dogs with mammary cancer and 50 control dogs and their concentrations were statistically related with these parameters. Significant difference was found between IL-8 serum levels in bitches with mammary neoplasia in relation to the control group and in relation to disease progression, the lymph node involvement and in relation to death. Low levels of IL-12 were observed in female dogs with tumors with a longer evolution, aged 10 years and in addition, the high survival rate was correlated with high levels of IL-12. Thus, increased IL- 8 and decreased IL-12 can be considered independent prognostic markers which confirm their role as tumorigenic and anti-tumorigenic cytokines, respectively. For the CA 15-3 were found higher levels in dogs with lymph node involvement and poor survival of the group. In estimating the number of copies of CANSINEs, significant difference was found regarding the parameters indicators of poor prognosis and low survival. Our results... (Complete abstract click electronic access below)

Submitted by GIOVANNA ROSSI VARALLO null (giovanna_vet03@yahoo.com.br) on 2016-12-26T14:11:39Z No. of bitstreams: 1 Tese Repositório.pdf: 4013000 bytes, checksum: 28bc5d1f53e09229a63ae5d00abb74ae (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-01-03T15:54:49Z (GMT) No. of bitstreams: 1 varallo_gr_dr_jabo.pdf: 4013000 bytes, checksum: 28bc5d1f53e09229a63ae5d00abb74ae (MD5) / Made available in DSpace on 2017-01-03T15:54:49Z (GMT). No. of bitstreams: 1 varallo_gr_dr_jabo.pdf: 4013000 bytes, checksum: 28bc5d1f53e09229a63ae5d00abb74ae (MD5) Previous issue date: 2016-11-16 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / O câncer de mama é uma doença heterogênea. Em mulheres está estabelecido um perfil imuno-histoquímico para utilização na rotina clínica de pacientes com câncer de mama. Os fenótipos luminal A, luminal B, HER2 superexpresso e triplo negativo apresentam diferenças prognósticas e preditivas. Entretanto, para a neoplasmas mamários em cadelas, não há uma classificação fenotípica definida. As diferenças prognósticas entre os fenótipos são atribuídas às assinaturas do tumor. Os estudos gênicos possibilitam averiguar o perfil genético da doença e a identificação de novos marcadores moleculares. O objetivo do estudo foi propor uma classificação fenotípica para a determinação de quatro fenótipos: luminal A, luminal B, HER2 superexpresso e triplo negativo, bem como avaliar a imunoexpressão dos marcadores anti-E-caderina, anti-claudinas 1, 3, 4, 7 e do anti-CD24 e anti-CD44. Na segunda etapa do estudo, objetivou-se avaliar o perfil de expressão gênica global dos carcinomas de mama e a diferença de expressão gênica entre os grupos fenotípicos. Para a avaliação imunohistoquímica, o grupo experimental foi composto por 110 amostras de câncer de mama dispostas em lâminas de tissue microarray. Obteve-se 42 tumores luminal A, 41 luminal B, 17 triplo negativo e 10 HER2 superexpresso. Constatou-se que estes últimos apresentam fenótipos mais agressivos e com menores sobrevidas. A expressão positiva da E-caderina foi mais acentuada nos grupos luminal A e luminal B, já a negativa no triplo negativo e no HER2 superexpresso. O padrão das expressões das claudinas 1, 3, 4 e 7 foi semelhante entre os subfenótipos. As células-tronco tumorais foram relacionadas aos graus histológicos mais agressivos (II e III). Para a avaliação da expressão gênica foram usadas 12 amostras de carcinoma mamário de cadelas e quatro mamas normais. Não foi evidenciada diferença na expressão gênica entre os grupos luminal A, luminal B, HER2 superexpresso e triplo negativo pelo microarranjo. O perfil gênico global das neoplasias estudadas mostrou 878 genes superexpressos e 821 genes subexpressos. A análise de enriquecimento aplicada aos genes superexpressos apontou o gene HYAL-1 como um potencial marcador diagnóstico e de recidiva. O PCR confirmou esse resultado. Conclui-se que a classificação molecular é uma ferramenta importante para a avaliação prognóstica dos pacientes com neoplasmas mamários. O gene HYAL-1 diferencialmente expresso participa da via de sinalização metabólica relacionada à carcinogênese mamária e a superexpressão está relacionada com o desenvolvimento do câncer e à recidiva. / The mammary cancer is a heterogeneous disease. In women, an immunohistochemical profile is established for use in the clinical routine of patients with breast cancer. The luminal A, luminal B, HER2 overexpressed and triple negative phenotypes present prognostic and predictive differences. However for mammary neoplasms in bitches, there is no definite phenotypic classification. Prognostic differences between phenotypes are attributed to tumor signatures. Genetic studies make it possible to ascertain the genetic profile of the disease and the identification of new molecular markers. The objective of the study was to propose a phenotypic classification for the determination of four phenotypes in bitches: luminal A, luminal B, HER2 overexpressed and triple negative, as well as to evaluate the immunoexpression of the anti-E-cadherin, anti-claudin markers 1, 3, 4, 7 and anti-CD24 and anti-CD44. In the second stage of the study, the objective was to evaluate the overall gene expression profile of mammary carcinomas and the difference in gene expression between the phenotypic groups. For the immunohistochemical evaluation, the experimental group consisted of 110 mammary cancer samples arranged in tissue microarray slides. 42 luminal A, 41 luminal B, 17 triple negative and 10 HER2 superexpressed tumors were obtained. It was verified that the latter present more aggressive phenotypes and with lower survival rates. Positive expression of E-cadherin was more pronounced in the luminal A and luminal B groups, whereas the negative expression in triple negative and HER2 overexpressed. The standard expression of claudins 1, 3, 4 and 7 was similar among subphenotypes. Tumor stem cells were related to the most aggressive histological grades (II and III). For the evaluation of gene expression, 12 samples of mammary carcinoma of bitches and four normal breasts were used. There was no difference in gene expression between the luminal A, luminal B, HER2 superexpressed and triple negative groups by the microarray. The overall gene profile of the neoplasms studied showed 878 overexpressed genes and 821 underexpressed genes. The enrichment analysis applied to the overexpressed genes pointed to the HYAL-1 gene as a potential diagnostic and relapse marker. PCR confirmed this result. It is concluded that molecular classification is an important tool for the prognostic evaluation of patients with breast neoplasms. The differentially expressed HYAL-1 gene participates in the metabolic signaling pathway related to mammary carcinogenesis and overexpression is related to the development of cancer and relapse. / CNPq: 140624/2013-9