Made available in DSpace on 2014-12-17T15:32:33Z (GMT). No. of bitstreams: 1 AntonioLAP.pdf: 801979 bytes, checksum: bfcf5910a257b5978efceb008f0c1016 (MD5) Previous issue date: 2006-09-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The unpredictable biologic behavior of the oral squamous cells carcinoma has determined extensive research on the evolution of such tumor. Due to the existing relation between the outer cell matrix and the tumor cells, the integrins have been used as markers in the predictive study of the cell behavior. This study aims to analyze immunohistochemically the expression of the integrin ?2?1, ?3?1, and ?5?1 connections for the collagen, the laminin and the fibronectin respectively in 15 cases of squamous cells carcinoma from the lower lip and 15 from the tongue, with different scores of malignance grading. A predominantly diffuse, cytoplasm and granular immunological marking was observed in the majority of the analyzed cases. According to the marking intensity, integrin ?2?1 appeared positive in 80% of the lip and in 93,3% of the tongue cases. The immunological reactivity of integrin ?3?1 was classified as positive in 60% of both the tongue and lip cases. For this integrin, 20% and 33.3% of the tongue and lip cases, respectively, were negative. In relation to integrin ?5?1 the intensity was classified as positive in 53,3% of the cases and strongly positive in 46,7% of those located in the lip. In the tongue carcinomas, the intensity was positive in 46,7% of the cases and strongly positive in 53,3%. The statistic analysis did not show any significant differences or correlation of expression between these integrins nor between the anatomical sites or between different scores of malignancy grading. The expressive immunological marking of the integrins, ?2?1, ?3?1, and ?5?1 in the studied cases of squamous cell carcinomas leads us to think of a great participation of these proteins in oral carcinogenesis; however, our results do not allow us to correlate its expression as an indicator of variations in the biological behavior of this neoplasia / A imprevisibilidade do comportamento biol?gico do carcinoma epiderm?ide oral vem justificando um grande n?mero de pesquisas utilizando biomarcadores que possam contribuir para um melhor entendimento do curso evolutivo dessa neoplasia. Por estarem envolvidas em rela??es entre as c?lulas tumorais e constituintes da matriz extracelular, as integrinas v?m sendo estudadas como poss?veis marcadores preditivos desse comportamento. Este estudo se prop?s a analisar, atrav?s do m?todo da imunohistoqu?mica, a express?o das integrinas ?2?1, ?3?1 e ?5?1, ligantes para o col?geno, laminina e fibronectina respectivamente, em 15 casos de carcinoma epiderm?ide de l?bio inferior e 15 de l?ngua, com diferentes escores de malignidade histol?gica. Observou-se uma imunomarca??o predominantemente difusa, citoplasm?tica e granular na maioria dos casos analisados. Quanto ? intensidade de marca??o, a integrina ?2?1 mostrou-se de forma positiva em 80% dos casos de l?bio e em 93,3% dos de l?ngua. A imunorreatividade da integrina ?3?1 foi classificada como positiva em 60% dos casos de l?bio e de l?ngua. Para esta integrina, 20% e 33,3% dos casos de l?bio e l?ngua, respectivamente, mostraram-se negativos. J? com rela??o ? integrina ?5?1a intensidade foi classificada como positiva em 53,3% dos casos e fortemente positiva em 46,7% daqueles localizados em l?bio. Nos carcinomas de l?ngua, a intensidade mostrou-se positiva em 46,7% dos casos e fortemente positiva em 53,3%. A an?lise estat?stica n?o demonstrou diferen?as nem correla??es significativas da express?o dessas integrinas nem entre os s?tios anat?micos, nem entre diferentes escores de grada??o histol?gica de malignidade. A expressiva imunomarca??o das integrinas ?2?1, ?3?1e ?5?1 nos casos de carcinomas epiderm?ides estudados nos leva a sugerir uma ampla participa??o dessas prote?nas na carcinog?nese oral; no entanto, nossos resultados n?o nos permitem correlacionar sua express?o como indicador de varia??es no comportamento biol?gico desta neoplasia

Les anatomopathologistes disposent d’outils permettant d’assister leurs décisions cliniques et d’évaluer les risques de récidive des patientes atteintes d’un cancer du sein. Parmi ceux-ci, le grade histologique du cancer du sein divise les patientes en trois sous-groupes pour lesquels le grade histologique 1 et 3 sont respectivement associés à de bons et mauvais pronostics. Cependant, cet outil est loin d’être parfait, dû au manque de reproductibilité de ce système et du risque de récurrence intermédiaire, peu informatif, des patients classés dans la catégorie « grade 2 ».<p>Afin de mieux caractériser ces tumeurs de risque intermédiaire, notre laboratoire a introduit un score appelé « Gene expression Grade Index (GGI) », basé sur l’expression de 97 gènes définis par microarrays. De façon intéressante, ce GGI permet de diviser les patientes de grade histologique 2, sur base de leur profil d’expression, en 2 groupes correspondant aux tumeurs de grade 1 ou aux tumeurs de grade 3. Cependant, bien que le GGI apporte une information importante, son applicabilité clinique est limitée par son prix et la nécessité d’utiliser du matériel congelé.<p>Durant ce travail de thèse, nous avons transposé la signature microarrays en un test RT-PCR, appelé PCR-GGI, basé sur l’expression de 8 gènes qui permet de reproduire les performances du GGI à partir de tissus congelés ou conservés dans de la paraffine. Cette amélioration permet de faciliter son utilisation en routine clinique. <p>De plus, nous avons approfondi notre connaissance du grade histologique, au niveau génomique et transcriptomique, et montré que les tumeurs mammaires (ER-positives) peuvent être divisées en deux groupes :un premier groupe de faible instabilité génomique, exprimant faiblement les gènes de prolifération et présentant un faible risque de récurrence ;et un deuxième groupe de haute instabilité génomique (impliquant principalement des amplifications localisées dans les régions 8q et 20q), une expression importante de gènes de prolifération et un mauvais pronostic.<p>D’autre part, les carcinomes canalaires in situ (DCIS) présentant des similarités avec les tumeurs invasives, nous avons voulu mieux comprendre le comportement du grade tumoral parmi ces tumeurs pré-invasives. Nous avons donc intégré le PCR-GGI au VNPI et défini le VNPI-GGI. Comparé au VNPI classique, le VNPI-GGI identifie mieux les patientes qui vont récidiver tôt dans les groupes de risque intermédiaire et haut, et permet donc d’éviter le sur-traitement.<p>Cependant, le calcul du VNPI est un travail fastidieux et le PCR-GGI seul ne permet pas de prédire les risques de récidives des DCIS. Nous avons donc cherché un nouveau marqueur pronostique. Alors, qu’il existe des preuves de plus en plus nombreuses supportant l’importance du rôle anti-tumoral des cellules myoépithéliales, nous avons montré qu’une diminution de l’expression de CD10 – un marqueur des cellules myoépithéliale – était hautement corrélée au risque de récidive. Ces résultats soulignent l’importance tant de l’agressivité de la tumeur que de son environnement directe, dans la progression tumorale.<p><p>En terme d’applications, les résultats obtenus durant ce travail de thèse nous ont permis de développer des outils utilisables par les cliniciens afin d’améliorer la prise en charge des patientes.<p><p><p><p>Traditional histopathological tools routinely used to evaluate breast cancer prognosis are designed to assist physicians in their evaluation of clinical outcome. The histological grade of invasive breast cancer, that assigns patients to one of 3 groups for which histological grade 1 and 3 tumors are respectively associated with lower and higher rate of recurrence, has long provided clinically important prognostic information. However, this tool is far from perfect due to concern over reproducibility and intermediate risk of recurrence of the histological grade 2 that is not informative for clinical decision. <p>To better characterize tumors classified as histological grade 2, our group has introduced a score called Gene expression Grade Index (GGI) based on a cassette of 97 genes defined by Microarrays. Interestingly, the GGI was able to reclassify patients with histological grade 2 tumors into 2 groups with distinct clinical outcomes similar to those of histological grade 1 and 3, respectively. However, its clinical applicability still remains expensive and often requires frozen tissue.<p>During this thesis work, we have transposed the GGI onto a qRT-PCR assay, called PCR-GGI, based on a set of 8 genes that could recapitulate in an accurate and reproducible manner the prognostic performance of GGI using both frozen and paraffin-embedded (FFPE) tumor samples, to facilitate its use in clinical practice. <p>Moreover, we have explored histological grade of invasive breast cancer at genomic and transcriptomic level and we have shown that two classes of ER-positive invasive breast cancer are observed: a first of low genomic instability, low proliferation gene expression and low risk of recurrence; and a second of high genomic instability (implying a major role for amplification of region located on chromosome arms 8q and 20q), high proliferation gene expression and worse prognosis.<p>In addition, since Ductal Carcinoma in situ (DCIS) and invasive breast cancer show concordant biologic behavior, we attempted to better understand the molecular basis of grade in pre-invasive breast cancer. We have then incorporated the PCR-GGI in the VNPI and defined the VNPI-GGI to improve its prognostic value. Compared to the classic VNPI, the VNPI-GGI had a better potential to identify early relapsing patients in the intermediate and high score group, and avoid under treatment in high-risk DCIS patients.<p>However, VNPI scoring is a tedious work and PCR-GGI alone can’t predict recurrence in pre-invasive breast cancer. We aimed then to find news prognosis marker in the field of DCIS. As there is now growing body of evidence supporting the role of myoepithelial cells (MECs) as natural tumor suppressors, we have showed that a decrease of CD10 expression- a surface biomarker of MECs – was significantly associated with an increased risk of relapse. <p>These results highlight the importance of assessing intrinsic DCIS properties as well as juxta-tumoral stroma, both seems to have a major role in DCIS progression.<p><p>In terms of applications, from these results obtained during this thesis work, we developed methods applicable into clinical practice to improve patients management.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

Made available in DSpace on 2014-12-17T15:32:18Z (GMT). No. of bitstreams: 1 ThaisGB.pdf: 4320063 bytes, checksum: c97c01facb2aed61dd49fc01568c6ca9 (MD5) Previous issue date: 2010-02-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The squamous cell carcinoma (SCC) is the most common malignant neoplasm of epithelial origin in oral cavity and present high capacity to invade adjacent structures. Traditionally, SCC has a predominance of 50 years male patients with long-time use of tobacco and alcohol, and the tongue is the most affected anatomic site. At present, there is an increasing incidence of SCC in patients below 40 years of age, who has been exposed or not to risk factors, mainly for tongue lesions. This study aims to analyze cell proliferation index using Ki-67 antigen in SCC of the tongue for two groups of different age range: until 40 years and older than 50 years. The first group was composed by 16 patients and the second one was composed by 20 patients. Clinicopathological features of the cases were also assessed. There was a male predominance in both groups. Tobacco and alcohol habits were common for patients until 40 years (72,2%), as well as for patients older than 50 years (52,9%). The first group had statistical association with the presence of regional metastases (p = 0,036) and with the most advanced stages of the disease (p = 0,012). Considering the histological malignancy grading, there was higher incidence (56,2%) of high malignancy grade tumors in the group of patients until 40 years old, but no statistical difference has found between groups and histologic malignancy grading. Regarding the immunohistochemical expression of Ki-67, there was no statistically significant difference between the antibody expression of the groups, as well as between other clinical and histopathological parameters. This study identified no significant difference regarding cell proliferation between the analyzed groups / O Carcinoma Epiderm?ide (CE) ? a neoplasia maligna de origem epitelial que mais ocorre na cavidade oral, com alta capacidade de invadir estruturas adjacentes. Classicamente, o CEO ocorre mais em homens com idade acima de 50 anos, que fazem uso de tabaco e ?lcool por longos per?odos de tempo, sendo a l?ngua, o s?tio anat?mico mais frequentemente acometido. Atualmente, vem se percebendo um aumento na incid?ncia dessa les?o em pacientes com idade abaixo dos 40 anos expostos ou n?o a fatores de risco, principalmente as les?es de l?ngua. O objetivo desta pesquisa foi analisar o ?ndice de prolifera??o celular, utilizando o anticorpo Ki-67em CEs de l?ngua em dois grupos de faixas et?rias distintas. Tamb?m, avaliaram-se as caracter?sticas cl?nico-patol?gicas dos casos constantes do estudo. A amostra se constituiu de 16 casos de pacientes com idade at? 40 anos e 20 casos de pacientes com idade acima de 50 anos. Em rela??o ?s caracter?sticas cl?nicopatol?gicas das les?es, o sexo masculino foi o mais acometido para os dois grupos, sendo evidenciado que o h?bito de beber e fumar foi frequente tanto para os pacientes com idade at? 40 anos (72,7%) como para os pacientes com idade acima dos 50 anos (52,9%). Foi poss?vel observar que houve uma associa??o estatisticamente significativa entre o grupo de pacientes com idade at? 40 anos e a presen?a de met?stase regional (p = 0,036), bem como entre o mesmo grupo e os est?gios mais avan?ados da doen?a (p = 0,012). Em rela??o ? grada??o histol?gica de malignidade, houve uma maior frequ?ncia de tumores classificados em alto grau de malignidade no grupo de pacientes com at? 40 anos (56,2%), mas n?o foi evidenciada diferen?a estat?stica entre os grupos e a grada??o histol?gica de malignidade. Quanto ? an?lise da express?o imuno-histoqu?mica pelo Ki-67, n?o houve diferen?a estatisticamente significativa entre a express?o do anticorpo para os grupos et?rios estudados nesta pesquisa, assim como n?o houve associa??o do ?ndice de positividade para o Ki-67 com os par?metros cl?nicos e histomorfol?gicos. Pode-se concluir que a prolifera??o celular n?o foi significativamente diferente entre os grupos que constitu?ram o presente estudo