Le travail présenté a porté sur l’analyse in vivo du proto-oncogène c-cbl, dont la forme connue est c-Cbl (p120cbl). Il s’agit d’une E3-Ubiquitine ligase et un poly-adaptateur moléculaire. Nous avons montré l’androgéno-dépendance de l’expression de c-Cbl dans les cellules germinales testiculaires et les cellules épithéliales de la prostate de rats et de souris. Nous avons montré la régulation anti-apoptotique exercée in vivo par la c-Cbl dans la prostate par comparaison des souris c-cbl invalidées ou non pour c-cbl (KO ou WT). L’effet exercé par c-cbl dans le testicule est pro-apoptotique (J.Cell Biol, 2005), que nous avons ultérieurement attribué à une nouvelle isoforme testiculaire de c-Cbl (Δ-c-Cbl). La comparaison des MEF KO et WT après induction d’apoptose par l’étoposide, a conforté l’effet anti-apoptotique exercée in vivo par c-Cbl dans la prostate. Elle a aussi montré la forte apoptose des MEF KO au peroxyde d’hydrogène : c-Cbl peut être considérée comme un protecteur du stress oxydant. L’intensité du stress oxydant associé aux cancers et leur forte résistance à l’apoptose sont des propriétés qui pourraient être reliées à c-Cbl. L’analyse in situ effectuée à partir de tumeurs congelées et de Tissue Microarrays (TMA) a montré une expression élevée de c-Cbl dans certains cancers, dont l’intensité pourrait correspondre à la gravité de l’atteinte anatomo-pathologique. La protéine c-Cbl est apparue être un marqueur d’agressivité du cancer de la prostate, probablement de l’ovaire, de l’utérus, du cerveau, du poumon, du colon et du rectum. Nous la considérons aussi comme une cible thérapeutique car, protecteur du stress oxydant, elle prendrait part à la résistance à l’apoptose des cellules tumorales. Un brevet a été déposé (2009, co-inventeurs : S.Yakoub et al). Un article rapportant ces résultats est en cours de soumission (S. Yakoub et al) / This work has focused on the in vivo analysis of the proto-oncogene c-cbl, coding for c-Cbl (p120cbl). We demonstrated the androgen-dependency of c-Cbl in the testicular germ cells and the prostatic epithelial cells of rats and mice. We then identified the anti-apoptotic regulation exerted by p120cbl in the prostate, comparing mouse c-cbl KO and WT, unlike this exerted in the testis (J.Cell Biol, 2005). We reported this difference to the high expression in testis of a new c-Cbl isoform, Δc-Cbl. The comparison of MEF KO and WT allowed confirming the anti-apoptotic regulation to etoposide exerted by c-Cbl. A very high apoptotic effect was observed in MEF KO with H2O2: c-Cbl is a strong stress oxidative protector. Knowing the intensity of oxidative stress in several cancers and their particular resistance to apoptosis as well, the in situ analysis of these malignancies was made from frozen tumours and tissue microassays (TMA). c-Cbl was indeed highly expressed and its intensity appears to reflect the aggressiveness of the pathology. c-Cbl could then be considered as a marker of severity of prostate cancer but probably also ovary, uterus, brain, lung, colon and rectum. It can also be considered as a therapeutic target involved in resistance to apoptosis as a stress oxidative protector. A patent was filed in the United States (2009, co-inventors: S. Yakoub et al)

Ce travail de thèse porte sur la quantification et la prise en compte des variations anatomiques en cours de radiothérapie guidée par l'image pour le cancer de la prostate. Nous proposons tout d'abord une approche basée population pour quantifier et analyser les incertitudes géométriques, notamment à travers des matrices de probabilité de présence de la cible en cours de traitement. Nous proposons ensuite une méthode d'optimisation des marges suivant des critères de couverture géométrique de la cible tumorale. Cette méthode permet d'obtenir des marges objectives associées aux différents types d'incertitudes géométriques et aux différentes modalités de repositionnement du patient. Dans un second temps, nous proposons une méthode d'estimation de la dose cumulée reçue localement par les tissus pendant un traitement de radiothérapie de la prostate. Cette méthode repose notamment sur une étape de recalage d'images de façon à estimer les déformations des organes entre les séances de traitement et la planification. Différentes méthodes de recalage sont proposées, suivant les informations disponibles (délinéations ou points homologues) pour contraindre la déformation estimée. De façon à évaluer les méthodes proposées au regard de l'objectif de cumul de dose, nous proposons ensuite la génération et l'utilisation d'un fantôme numérique reposant sur un modèle biomécanique des organes considérés. Les résultats de l'approche sont présentés sur ce fantôme numérique et sur données réelles. Nous montrons ainsi que l'apport de contraintes géométriques permet d'améliorer significativement la précision du cumul et que la méthode reposant sur la sélection de contraintes ponctuelles présente un bon compromis entre niveau d'interaction et précision du résultat. Enfin, nous abordons la question de l'analyse de données de populations de patients dans le but de mieux comprendre les relations entre dose délivrée localement et effets cliniques. Grâce au recalage déformable d'une population de patients sur une référence anatomique, les régions dont la dose est significativement liée aux événements de récidive sont identifiées. Il s'agit d'une étude exploratoire visant à terme à mieux exploiter l'information portée par l'intégralité de la distribution de dose, et ce en fonction du profil du cancer. / This work deals with the quantification and the compensation of anatomical deformations during image-guided radiotherapy of prostate cancer. Firstly, we propose a population-based approach to quantify the geometrical uncertainties by means of coverage probability matrices of the target tumor during the treatment. We then propose a margins optimization method based on geometrical coverage criteria of the tumor target. This method provides rationnal margins models associated to the different geometrical uncertainties and patient repositioning protocols. Secondly, we propose a method to estimate the locally accumulated dose during the treatment. This method relies on a deformable image registration process in order to estimate the organ deformations between each treatment fraction and the planning. Different registration methods are proposed, using different level of user interactions (landmarks specification or delineations) to constrain the deformation estimation. In order to evaluate the performance of the proposed methods, we then describe the generation of a numerical phantom based on a biomechanical model. The results are presented for the numerical phantom and real clinical cases. We show that the benefit brought by the manual placement of some landmarks to constrain the registration represents a good compromise between the required interaction level and the dose estimation accuracy. Finally, we address the issue of the analysis of population data in order to better understand the relationship between the locally delivered dose and clinical effects. With deformable image registration of a population of patients on an anatomical template, regions whose dose is significantly associated with recurrence events are identified. This last part is an exploratory study aiming to better use the information carried by the entire distribution dose, and according to the cancer profile.

Le cancer de la prostate représente la deuxième cause de décès liée au cancer. Des stratégies thérapeutiques ciblant des mécanismes moléculaires conduisant à la résistance doivent donc être développées. Une stratégie visant à améliorer les traitements du cancer de la prostate consiste à cibler les gènes qui sont activés lors de la disparition des androgènes, soit pour retarder ou empêcher l'émergence du phénotype de résistance à la castration. Le but de cette thèse est d'identifier et de développer des petites molécules inhibitrices ciblant des interactions protéine-protéine impliquées dans le cancer de la prostate. Cette thèse porte sur l'étude de deux protéines cruciales liées au cancer de la prostate, à savoir, la protéine de choc thermique de bas poids moléculaire (Hsp27) et la protéine TCTP. Nous avons validé deux composés ciblant TCTP en utilisant une chimiothèque dédiée à l'inhibition d'interaction protéine-protéine. Des tests fonctionnels sont actuellement mis au point pour évaluer la capacité de ces molécules à être proposées comme composés potentiels contre le cancer de la prostate. / Prostate Cancer (PCa) is one of most common malignancies, being the second leading cause among cancer-related death. Additional therapeutic strategies targeting molecular mechanisms mediating resistance must be developed because of the defects of docetaxel-based treatments. One strategy to improve therapies in advanced PCa involves targeting genes that are activated by androgen withdrawal, either to delay or prevent the emergence of the CR phenotype. The purpose of my thesis is to identify & develop small molecules inhibitors targeting PPIs involved in prostate cancer. we focuses on 2 crucial prostate cancer related proteins, namely, the small molecular weight Heat shock protein 27 (Hsp27) and the Translationally Controlled Tumor Protein (TCTP). We have validated 2 compounds targeting TCTP by using a "PPI Inhibitor-like" dedicated chemical library. Functional tests are now being developed to evaluate the capacity of such molecules to be proposed as potential compounds against prostate cancer.

No description available.

OBJETIVO: Avaliar a relação entre mensurações de gordura (subcutânea e periprostática) realizadas em ressonância magnética, com o prognóstico do paciente portador de câncer de próstata. MATERIAIS E MÉTODOS: A mensuração da gordura periprostática e subcutânea com exame de ressonância magnética com ênfase na próstata foram realizadas em 58 pacientes com o diagnóstico histopatológico de câncer de próstata. Dados demográficos, clínicos e patológicos dos pacientes foram coletados e a correlação com o prognóstico, realizada. RESULTADOS: Na análise univariada, as gorduras periprostática e a subcutânea indicaram serem preditores de evolução desfavorável com significância estatística para o observador 1, no caso da gordura periprostática e para o observador 2 no caso da gordura subcutânea. Na análise multivariada, não houve associação com significância estatística. CONCLUSÃO: A associação entre obesidade e o câncer de próstata é complexa. Os dados obtidos neste estudo indicam que a medida da gordura periprostática, pelas imagens em T2, na RM, podem ser um preditor independente, da evolução desfavorável de pacientes com neoplasia de próstata / PURPOSE: Evaluate the relationship between fat measurements (subcutaneous and periprostatic) performed on MRI, with the prognosis of the patient with prostate cancer. MATERIALS AND METHODS: Measurements of periprostatic and subcutaneous fat with magnetic resonance imaging with emphasis on the prostate were performed in 58 patients with the histopathological diagnosis of prostate cancer. Demographic, clinical and pathological data of the patients were collected and the correlation with the prognosis was performed. RESULTS: In the univariate analysis, the periprostatic and subcutaneous fat were predictors of unfavorable evolution with statistical significance for the observer 1, in the case of the periprostatic fat and for the observer 2 in the case of the subcutaneous fat. In the multivariate analysis, there was no association with statistical significance. CONCLUSION: The association between obesity and prostate cancer is complex. The data obtained in this study indicate that the measurement of periprostatic fat by T2-weighted images in MRI may be an independent predictor of the unfavorable evolution of patients with prostate neoplasia

Cette thèse concerne le développement et l'évaluation des nanoparticules pour le traitement du cancer et plus particulièrement pour le cancer de la prostate.Le manuscrit comprend une revue de la littérature sur l'application de la nano médecine pour le traitement du cancer de la prostate. Dans la première partie expérimentale, des nanoparticules d'or fonctionnalisées ont été caractérisées et chargées avec le docétaxel par adsorption non covalente.Ces nanoparticules d'or ont montré un effet cytotoxique in vitro prolongé contre les cellules cancéreuses de la prostate. La deuxième partie expérimentale de cette thèse décrit une étude de synthèse et une nano-précipitation de polyesters pour la co-délivrance de deux médicaments chimio-thérapeutiques, le docétaxel (DOC) et la mitoxantrone (MIT). Les polycaprolactone, poly(acide lactique) et poly(lactide-co-glycolide) ont été synthétisés par polymérisation par ouverture de cycle avec des poids moléculaires différents de polyéthylène glycol. Des nanoparticules monodisperses d’un diamètre d’environ 80 nm ont été obtenues et se sont avérées être efficaces contre les cellules cancéreuses de la prostate quand cela est chargé en MIT et DOC. De plus, un effet synergique a été observé en utilisant des combinaisons de ces nanoparticules. Par conséquent, ces nanoparticules, à base de polyester, ont de potentielles applications cliniques. / This thesis concerns the development and evaluation of nanoparticles for cancer treatment, and in particular to prostate cancer. The manuscript includes a literature review on the application of nanomedicine to the treatment of prostate cancer. In the first experimental part, functionalized gold nanoparticles were characterized and loaded with docetaxel by non covalent adsorption. These gold nanoparticles showed a sustained cytotoxic effect in vitro against prostate cancer cells. The second experimental part of this thesis describes a study of synthesis and nanoprecipitation of polyesters for the co-delivery of two chemotherapeutic drugs, docetaxel (DOC) and mitoxantrone (MIT). Polycaprolactone, poly(lactic acid) and poly (lactide-co-glycolide) were synthesized by ring-opening polymerization with different molecular weights of polyethylene glycol. Monodisperse nanoparticles with diameters of less than 80 nm were produced and were shown to be effective against prostate cancer cells when loaded with MIT and DOC. Moreover, a synergistic effect was observed using combinations of these nanoparticles. Therefore, these polyester based nanoparticles have potential clinical applications.

O câncer de próstata (CaP) é a segunda causa mais comum de câncer em homens. De acordo com o INCA, no Brasil, em 2016, estimam-se aproximadamente 61.200 novos casos de câncer de próstata. Objetivo: Identificar as características demográficas e epidemiológicas, bem como dados do estadiamento tumoral dos pacientes com CaP atendidos na Unidade de Oncologia do Hospital Marcio Cunha na cidade de Ipatinga-MG nos anos de 2012, 2013 e 2014. Metodologia: Trata-se de um estudo retrospectivo e descritivo onde foram analisados 668 prontuários de pacientes, com registro do diagnóstico anatomopatológico, atendidos nos anos de 2012, 2013 e 2014, conforme lista fornecida pela instituição, com diagnóstico de CaP cadastrados com CID-10 - C 61. As variáveis analisadas foram: procedência, ano do diagnóstico, faixa etária, raça autodeclarada, fatores de risco como tabagismo, etilismo, história familiar de CaP, PSA total ao diagnóstico, tipo histológico da biópsia, score de Gleason da biópsia, tipo histológico da peça cirúrgica, score de Gleason da peça cirúrgica. Os dados foram analisados empregando-se estatística descritiva e inferencial, utilizando o software SPSS, versão 19.0. Resultados: A maior incidência de casos de CaP foram provenientes das cidades mais populosas da microrregião de saúde analisada e faixa etária mais prevalente foi entre 61 e 80 anos com prevalência em pardos e brancos e com histórico familiar de 17,2% de parentes de primeiro grau; com o pai em 37,3%, o irmão em 60,8% e filho em 1,9%. Apenas 165 (25,9 %) eram fumantes e 20,8% etilistas. Os níveis de PSA ficaram entre 4,1ng/ e 10ng/ml (49,5%) e quanto maior a faixa etária maiores os valores do PSA. Pacientes pardos apresentaram PSA total mais elevado. Ao avaliarmos se existia relação entre os níveis de PSA total com fatores de risco como tabagismo, etilismo e histórico familiar, somente houve relação estatisticamente significativa com o etilismo. Houve concordância do score de Gleason entre biópsia e peça cirúrgica em 70%, subgraduação em 18,7% e supergraduação em 11,3%. Comparando a idade dos pacientes com Score de Gleason, quanto maior a idade do paciente maior foi o Score de Gleason do material obtido pela biópsia via transretal Pacientes tabagistas e etilistas apresentaram Score de Gleason da peça cirúrgica mais elevados. Conclusão: A concordância entre o Score de Gleason da biópsia e o Score de Gleason da peça cirúrgica foi de 70%; etilistas apresentaram PSA mais elevados; quanto maior foi a faixa etária, mais indiferenciado foi o tumor ( biópsia); pacientes tabagistas e etilistas apresentaram tumores mais indiferenciados na peça cirúrgica; este é o primeiro estudo epidemiológico de CaP desenvolvido na região do Vale do Aço, a caracterização sócio demográfica e as associações aqui encontradas podem contribuir com programas para desenvolver ações de controle do CaP nesta região. / Prostate cancer (PCa) is the second most common cause of cancer in men. According to INCA, in Brazil, in 2016, approximately 61,200 new cases of prostate cancer are estimated. Objective: To identify the demographic and epidemiological characteristics, as well as data on the tumor staging of patients with PCa treated at the Oncology Unit of Hospital Marcio Cunha in the city of Ipatinga-MG in the years of 2012, 2013 and 2014. Methodology: This is a retrospective and descriptive study where 668 patients\' records, with a diagnosis of pathological diagnosis, were analyzed in the years 2012, 2013 and 2014, according to the list provided by the institution, with a diagnosis of PCa registered with ICD-10-C 61. The analyzed variables were: origin, year of diagnosis, age group, self-reported race, risk factors such as smoking, alcoholism, family history of PCa, total PSA at diagnosis, histological type of biopsy, Gleason score of biopsy, histological type of the surgical specimen, Gleason score of the surgical specimen. Data were analyzed using descriptive and inferential statistics, using SPSS software, version 19.0. Results: The highest incidence of PCa cases came from the most populated cities of the analyzed health micro-region and the most prevalent age group was between 61 and 80 years old, with prevalence in brown and whites and with a family history of 17.2% of first-degree relatives degree; With father in 37.3%, brother in 60.8% and son in 1.9%. Only 165 (25.9%) were smokers and 20.8% were alcoholics. PSA levels ranged from 4.1ng / e to 10ng / ml (49.5%) and the higher the age group the higher the PSA values. Brown patients had higher total PSA. When we evaluated whether there was a relationship between total PSA levels and risk factors such as smoking, alcohol consumption and family history, there was only a statistically significant relationship with alcohol consumption. There was concordance of the Gleason score between biopsy and surgical specimen in 70%, subgrade in 18.7% and overdose in 11.3%. Comparing the age of patients with Gleason score, the greater the patient\'s age, the greater the Gleason score of the material obtained by the transrectal biopsy. Smokers and alcoholists presented the highest Gleason score of the surgical specimen. Conclusion: The agreement between the Gleason score of the biopsy and the Gleason score of the surgical specimen was 70%; Higher PSA levels; The longer the age group, the more undifferentiated was the tumor (biopsy); Smokers and alcoholics presented more undifferentiated tumors in the surgical specimen; This is the first epidemiological study of PCa developed in the Vale do Aço region, the socio-demographic characterization and the associations found here can contribute with programs to develop actions of control of PCa in this region.

Die vorliegende Habilitationsschrift "Neue Serummarker bei urologischen Malignomen mit dem Schwerpunkt Prostatakarzinom und Anwendung von Proteinase-Inhibitoren in der Therapie des Prostatakarzinoms" faßt Ergebnisse zusammen, die ich in den Jahren von 1996 bis 2000 als Erstautor in wissenschaftlichen Artikeln von peer reviewed Zeitschriften veröffentlicht habe. Zusätzlich werden 14 Arbeiten mit ihren Aussagen eingeschlossen, bei denen ich als Koautor beteiligt war. Gegenstand der Habilitationsschrift sind Untersuchungen zur diagnostischen Optimierung des Tumormarkers Prostataspezifisches Antigen (PSA) und zum Expressionsverhalten von CD44-Proteinen bzw. Matrix-Metalloproteinasen (MMPs) als potentielle, neue Marker bei urologischen Karzinomen. Die nachgewiesene Bedeutung der MMPs bei der Tumorprogression und -metastasierung hat mich dazu veranlaßt, tierexperimentelle Studien zur Hemmung der MMP-Aktivität mit synthetischen Inhibitoren in meine Arbeit aufzunehmen. Zielstellung war hierbei die Evaluierung neuer Therapieoptionen beim fortgeschrittenen Tumor. Im Mittelpunkt der Untersuchungen steht das Prostatakarzinom (PCa) als häufigster maligner Tumor des Mannes. 1. Das PSA ist ohne Zweifel der beste Tumormarker in der Diagnostik des PCa. Der Optimierung dieses Markers wird große Bedeutung zugemessen. Dabei werden verschiedene Konzepte verfolgt, wobei die Bestimmung der Isoformen des PSA die zur Zeit erfolgreichste Richtung zu sein scheint. Die Ergebnisse meiner u.a. im Rahmen von Multizenterstudien durchgeführten Untersuchungen belegen den diagnostischen Nutzen der zusätzlichen Bestimmung des f-PSA%, um PCa-Patienten früher zu erkennen und besser gegenüber Patienten mit benigner Prostatahyperplasie (BPH) abgrenzen zu können. Ein weiteres Ergebnis der Untersuchungen zur diagnostischen Validität anderer PSA-Isoformen ist die Feststellung, daß die Bestimmung des gebundenen PSA keinen Vorteil gegenüber dem f-PSA% hat. Widersprüchliche Angaben in der Literatur konnten damit ausgeräumt werden. Diese Ergebnisse veranlaßten die Firma Roche als Kooperationspartner, die Weiterentwicklung eines ACT-PSA Prototyp-Testsystems einzustellen. Die Ergebnisse meiner Untersuchungen zu den PSA-Isoformen haben inzwischen Eingang in den klinischen Alltag gefunden und werden in der Urologischen Klinik der Charité genutzt. Es wurden Entscheidungsgrenzen für den f-PSA%-Wert zur Indikationsstellung von Stanzbiopsien der Prostata als Klinikstandard erarbeitet. 2. Bei verschiedenen urologischen Tumoren wurde das Expressionsverhalten von CD44-Proteinen und MMPs bzw. deren Inhibitoren bestimmt, um eine mögliche Bedeutung bei der Tumorprogression zu erfassen. Diese Untersuchungen sind gleichzeitig Voraussetzung für eine mögliche Anwendung der Komponenten in der Diagnostik und Therapiekontrolle bei diesen Tumorentitäten. Im Gegensatz zu anderen menschlichen Tumoren konnten bei den untersuchten urologischen Tumoren keine veränderten Serumkonzentrationen der CD44-Proteine einschließlich der Varianten nachgewiesen werden. Daher habe ich weiterführende Studien zu den CD44-Proteinen nicht durchgeführt. 3. Im Tumorgewebe sowie im Plasma von Patienten mit PCa und Nierenzellkarzinom konnte ich signifikante Veränderungen von MMPs und deren Inhibitoren nachweisen. Die Situation im Gewebe spiegelt sich zum Teil im Blut der Patienten wider. Diese Beobachtungen beweisen die Bedeutung der MMPs bei der Tumorprogression und -metastasierung. Prinzipiell kann die Bestimmung einzelner MMPs bzw. TIMPs in der Diagnostik von urologischen Tumoren genutzt werden. Die niedrige Sensitivität in der individuellen Erfassung des einzelnen Tumorpatienten schränkt jedoch die praktische Anwendung ein. 4. Die veränderte MMP-Expression bzw. die Dysbalance zwischen MMPs und TIMPs hat mich veranlaßt, synthetische Inhibitoren zur Blockierung der MMP-Aktivität in einem Standardtiermodell des menschlichen PCa einzusetzen. In einer ersten Untersuchung am Dunning-Tumor der Ratte wurde nachgewiesen, daß dieser Tumor MMP9 exprimiert und die Serumkonzentration mit der Tumorgröße korreliert. In weiteren tierexperimentellen Untersuchungen wurde der Einfluß von Batimastat, einem Breitspektrum-Inhibitor der MMPs und einem neuentwickelten, selektiveren Inhibitor (Icol) auf das orthotope Tumorwachstum ermittelt. Beide Substanzen führten zu einer Hemmung des lokalen Tumorwachstums. Durch Applikation von Icol wurde eine Reduzierung des Tumorgewichtes um 90% im Vergleich zu den unbehandelten Kontrolltieren erreicht. Diese Beobachtungen haben eine doppelte Bedeutung. Zum einen beweist die Hemmwirkung von synthetischen MMP-Inhibitoren im Tiermodell die Funktion der MMPs bei der lokalen Tumorprogression. Zum anderen werden durch diese erfolgreichen tierexperimentellen Studien mit neuen Substanzen Voraussetzungen für die klinische Anwendung bei Patienten mit hormonrefraktärem PCa geschaffen. / The aim of my "habilitation thesis" was to evaluate the diagnostic validity of prostate-specific antigen (PSA) in serum and tissue, the serum pattern of CD44 proteins and of the matrix metalloproteinases (MMPs) in serum and tissue of urological malignancies. As MMPs seem to play an important role in tumor progression and metastasis, animal studies were additionally initiated in order to investigate the influence of synthetic inhibitors of MMPs on prostate cancer. 1. PSA is the most important and accurate tumor marker in prostate cancer diagnosis. However, PSA is an organ-specific marker, but is not tumor-specific. Elevated PSA concentrations are seen with non-malignant prostatic diseases like benign prostatic hyperplasia (BPH). Moreover, not all patients with prostate cancer have elevated PSA concentrations. In order to optimize the diagnostic validity of PSA, several concepts have been developed. Determination of the PSA isoforms in serum could help discriminate between prostate cancer and BPH. In various own studies, including a multicenter clinical trial, the determination of free PSA and the calculation the ratio of free PSA to total PSA (fPSA/tPSA) has proven to be a promising tool in prostate cancer diagnosis. Regarding the diagnostic validity of the complexed PSA conflicting data exist. Our results, using a newly developed alpha-1-antichymotrypsin-PSA (ACT-PSA) assay by Roche are contradictory to recent published data. Based on data of a multicenter trial, the determination of ACT-PSA as well as the ACT-PSA to tPSA ratio did not improve the differential diagnostic impact in patients undergoing evaluation for prostate cancer compared to the ratio fPSA/tPSA. 2. In various malignant diseases characteristic alterations in the expression of CD44 proteins and their variants have been observed. In contrast to those observations in other carcinomas, the determination of soluble CD44 proteins in serum is not suitable for detecting and staging patients with urological malignant tumors. Therefore, further investigation have not been performed. 3. Matrix-metalloproteinases (MMP) form a group of endogenous proteases with the common ability to degrade various components of the extracellular matrix. It could be demonstrated that increased levels of MMP are associated with the invasive and metastatic potential in human malignant tumors. However, little is known about the role of MMPs in renal cell carcinoma. In own study significant changes of MMP expression have been observed. Although changes in specific MMPs might be characteristic for renal carcinoma tissues and might be partly reflected in the blood, data shown that even MMP-9 as the best plasma marker, had a low sensitivity in detecting renal cell carcinoma. Increased concentrations of MMP-9 in tumor tissue may have important implications for the therapeutic potential of synthetic inhibitors of MMPs. 4. The importance of inhibitors of MMPs in cancer has been demonstrated in various studies. In own investigations, altered levels of MMPs and their specific inhibitors have been elucidated in prostate cancer. Therefore, a study to evaluate the efficacy of synthetic MMP inhibitors (batimastat, Icol) in a standard prostate cancer animal model was performed. Previously, the high expression of MMP-9 in this prostate cancer (Dunning tumor) compared with normal prostatic tissue could be demonstrated. Batimastat and the newly developed inhibitor Icol reduced the orthotopic tumor weights up to 90% in a dose-dependent manner. This results confirmed the importance of MMPs and their inhibitors in tumor progression. It can be concluded that selective inhibition of MMP activity is a novel therapeutic approach, which bears promise for studies in patients with hormone-refractory prostate cancer.

Introdução: O câncer de próstata (CaP) é o tumor mais freqüente do homem no Brasil tendo sido estimados mais de 52.350 novos casos em 2010, sendo a segunda causa de óbito por câncer em homens. O prognóstico depende fundamentalmente dos níveis séricos de Prostatic Specific Antigen (PSA) estádio tumoral (TNM) e grau de diferenciação histológica (Gleason). Porém esses têm sido insuficientes na definição do prognóstico da neoplasia. Por isso pesquisas têm sido direcionadas para a identificação de alterações moleculares que possam prever o potencial de agressividade do câncer de próstata. Metaloproteinases da matriz (MMP) são proteínas pertencentes a uma família de aproximadamente 30 enzimas proteolíticas ou endoproteinases que degradam vários componentes da matriz extracelular. A detecção de sua expressão tem sido estudada como marcador sensível e específico de vários tumores, principalmente as MMP pertencentes ao grupo das gelatinases MMP-2 e MMP-9. Objetivo: o objetivo deste nosso trabalho foi avaliarmos pela técnica de qRT-PCR e imuno-histoquímica os níveis de expressão dos genes das MMP pertencentes ao grupo das gelatinases, MMP-2 e MMP-9, bem como outros sabidamente envolvidos em suas vias de ativação (MMP-14, IL-8) e inibição (TIMP-1, TIMP-2, RECK e TGF-) no câncer localizado de próstata. Material e Métodos: O estudo consistiu na análise de espécimes de 79 pacientes com câncer da próstata submetidos a prostatectomia radical entre setembro de 1997 e fevereiro de 2000. Esses oito genes foram então testados quanto a seu valor prognóstico no câncer da próstata através da técnica de reação em cadeia da polimerase quantitativa com transcriptase reversa (qRT-PCR). Análise proteica foi feita a partir de 40 pacientes deste pool. O grupo controle foi composto de tecido de 11 pacientes com hiperplasia benigna da próstata (HPB) tratados cirurgicamente com prostatectomia retropúbica. Resultados: MMP-9 esteve superexpressa e MMP-2, TIMP-1, TIMP-2, MMP 14, IL-8, TGF- e RECK se mostraram subexpressos em tecido representativo de CaP quando comparado com HPB. A análise dos níveis de expressão dos genes com o escore de Gleason, mostrou que MMP-2 e TIMP-2 mesmo mantendo-se subexpressos, tiveram uma expressão maior entre os pacientes que apresentavam Gleason 7 (p=0,04 e p=0,02 respectivamente). De acordo com o valor de PSA préoperatório, encontramos diferenças na expressão de MMP-9. Pacientes que apresentavam um PSA pré-operatório 10 ng/mL possuíam uma mediana de expressão maior que aqueles cujo PSA pré-operatório <10 ng/mL com medianas de expressão de 5,62 e 2,76 respectivamente (p=0,033). Não encontramos diferenças estatísticas entre pacientes que apresentavam ou não recidiva bioquímica quanto a expressão dos 8 genes estudados. Porém o gene da MMP-9 apresentou uma diferença estatística marginal apresentando uma mediana de expressão de 6,29x nos pacientes que apresentaram recidiva bioquímica e de 3,25 nos pacientes que não apresentaram recidiva bioquímica (p=0,090). De acordo com a expressão proteica, encontramos uma maior positividade em MMP-9, MMP-2, TGF-, IL-8 e MMP-14. De acordo com os fatores prognósticos encontramos associação de TIMP-1 com recidiva bioquímica. Conclusão: Encontramos uma superexpressão de MMP-9 e uma subexpressão de MMP-2, TIMP-1, TIMP-2, MMP-14, RECK, IL-8 e TGF- no CaP. Considerando os fatores prognósticos encontramos que aumentados níveis de expressão do gene da MMP-9 associou-se a aumentados níveis de PSA, e mostrou uma tendência de associação com recidiva bioquímica. De acordo com a expressão proteica encontramos que a ausência de TIMP-1 pode ser um indicativo de recidiva bioquímica / Introduction: Currently, Prostate cancer (PCa) is the most common tumor in men in Brazil. It was estimated that more than 52,350 new cases were diagnosed in 2010, being the second cause of death by cancer in man. The prognosis depends mainly on Prostate Specific Antigen (PSA) serum levels, tumor stage (TNM) and histological grade (Gleason), but these parameters, even combined, are insufficient to define the correct prognosis of PCa. Therefore research has been directed towards the identification of molecular alterations that may predict potential aggressiveness of PCa. Matrix metalloproteinases (MMPs) are proteins that belong to a family of about 30 proteolytic enzymes that degrade various components of the extracellular matrix. The analysis of MMPs expression has been studied as a sensitive and specific marker of prognosis of several tumors, and special attention was focused in the group of gelatinases, MMP-2 and MMP-9.Objective: The aim of this study was to evaluate the expression levels of MMP-2 and MMP-9 genes and proteins by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry in localized PCa. We also evaluated the expression of genes that are involved in the control of MMP-2 and MMP as activators (MMP- 14, IL-8) or inhibitors (TIMP-1, TIMP-2, RECK and TGF-).Materials and Methods: The casuistic consisted of 79 surgical specimens from patients with localized PCa who underwent radical prostatectomy between September 1997 and February 2000. The control group was composed of specimens from 11 patients with benign prostatic hyperplasia (BPH) treated surgically with retropubic prostatectomy. The results of the 8 genes expression, through qRTPCR and immunohistochemistry, were correlated to the diagnosis and prognosis of PCa. The protein expression analysis was carried out in 40 patients of the casuistic. Results: The MMP-9 was overexpressed, while MMP-2, TIMP-1, TIMP-2, MMP-14, RECK, IL-8, and TGF- were underexpressed in malignant prostate tissue compared to BPH. Patients with Gleason7 had higher expression of MMP-2 and TIMP-2 (p=0.04, p=0.02 respectively). According to the preoperative PSA value, we found that patients with preoperative PSA10 ng/mL had a median of expression of 5.62 compared to 2.76 when PSA<10 ng/mL (p=0.033). There were no statistical differences between expression of the eight genes and biochemical recurrence during follow up. However, the higher MMP-9 expression was marginally associated with recurrence, the median was of 6.29 in recurrence patients compared to 3.25 in those without recurrence (p=0.090). Regarding the protein expression, we found a higher positivity of MMP-9, MMP-2, TGF-, IL-8 e MMP-14 expression in PCa, and a correlation between the lack of TIMP-1 and tumor recurrence. Conclusion: MMP-9 is overexpressed while MMP-2, TIMP-1, TIMP-2, MMP-14, RECK, TGF- and IL-8 are underexpressed in CaP. According to the prognostic factors, we observed that increased level of MMP-9 was associated with pre-surgical PSA10 ng/mL. Also there was a tendency of association between higher MMP- 9 expression and biochemical recurrence. Overexpression of MMP-9 can be explained by the underexpression of their major inhibitors TIMP-1 and RECK. According to protein expression we found that absence of TIMP-1 is correlated with biochemical recurrence in the PCa

This study determined the impact of vitamin C dose on genistein-induced apoptosis in LNCaP cancer cells at various treatment regimens in vitro. Although the linear regression of viability assay (MTT) indicated a p-value = 0.11; NBT assay reveal a declining SOD activity during cell death. Apoptosis induction was the main mode of treatment induced cell death. The overall data showed the trend of treatment efficacy as;(Gen 10uM + Vit C 40uM) > (Gen 30uM + Vit C 40uM) > (Gen 70uM + Vit C 40uM) > 10uM genistein > 70uM genistein. The chi-square test for comparing necrosis, apoptosis and life cells showed that Vitamin C could impact genistein-induced apoptosis in LNCaP cells (p = 0.0003). This study forms the basis for in vivo studies of the impact of vitamin C on genistein-induced apoptosis in LNCaP prostate cancer cells. / Includes bibliography. / Thesis (M.S.)--Florida Atlantic University, 2015. / FAU Electronic Theses and Dissertations Collection