No / Dystroglycan is a ubiquitously expressed cell adhesion molecule frequently found to be altered or reduced in adenocarcinomas, however the mechanisms or consequences of dystroglycan loss have not been studied extensively. We examined the consequence of overexpression or RNAi depletion of dystroglycan on properties of in vitro growth migration and invasion of LNCaP, PC3, and DU145 prostate cancer cell lines. RESULTS: Using LNCaP cells we observed cell density-dependent changes in beta-dystroglycan with the appearance of several lower molecular weight species ranging in size from 43 to 26 kDa. The bands of 31 and 26 kDa were attributed to proteolysis, whereas bands between 43 and 38 kDa were a consequence of mis-glycosylation. The localization of beta-dystroglycan in LNCaP colonies in culture also varied, cells with a mesenchymal appearance at the periphery of the colony had more pronounced membrane localization of dystroglycan. Whereas some cells demonstrated nuclear dystroglycan. Increased dystroglycan levels were inhibitory to growth in soft agar but promoted Matrigel invasion, whereas reduced dystroglycan levels promoted growth in soft agar but inhibited invasion. Similar results were also obtained for PC3 and DU145 cells. This study suggests that changes in beta-dystroglycan distribution within the cell and/or the loss of dystroglycan during tumorigenesis, through a combination of proteolysis and altered glycosylation, leads to an increased ability to grow in an anchorage independent manner, however dystroglycan may need to be re-expressed for cell invasion and metastasis to occur.

<p>Adenokarcinom prostate (PCa) je najče&scaron;ći karcinom u mu&scaron;karaca. Intraepitelne prostatične neoplazme visokog gradusa (HGPIN) su lezije koje prethode nastanku invazivnog karcinoma i podrazumevaju kompletno odsustvo bazalnih ćelija i invaziju strome malignim acinusima. Estrogeni receptor &beta; (ER&beta;) se nalazi u jedrima bazalnih i sekretornih ćelija acinusa i delimično u stromalnim ćelijama. Cilj istraživanja je da prikaže i lokalizuje ER&beta; u različitim morfolo&scaron;kim lezijama prostate: hiperplaziji (BHP), PINu i u PCa sa različitim Gleason scorom. Pretpostavlja se da prekancerozne lezije u svojim različitim fazama evolucije ne koreliraju u potpunosti sa ekspresijom ER&beta;. LGPIN pokazuje ekspresiju, dok u HGPINu nema ekspresije. Takođe je pretpostavka da ekspresija ER&beta; postoji u većine srednje diferentovanih PCa, te da se ekspresija posmatranog receptora gubi sa porastom Gleason scora. Ispitivano je pet grupa bolesnika:&nbsp; kontrolna grupa sa BHP i četiri eksperimentalne grupe (PIN i 3 različite grupe PCa). Studija je sprovedena na mu&scaron;karcima različite starosti u periodu 2010&ndash;2012. Nijedan pacijent nije prethodno primio hormonsku terapiju. Sekstant biopsije prostate su bojene na ER&beta; (Novocastra). Lokalizacija i intenzitet ER&beta; ekspresije prikazani su kroz skor: 0 = nula; 1 = &lt;1%; 2 = 1&ndash;10%; 3 = 11&ndash;33%; 4 = 34&ndash;66%; 5 = &gt; 66%. Pozitivni fibroblasti i endotelne ćelije su kori&scaron;ćene za poređenje. Smanjena ekspresija ER&beta; primećena je kod malignih i premalignih lezija prostate naspram BHP. Ekspresija ER&beta; u epitelnim ćelijama acinusa bila je najslabija u dobro diferentovanim PCa. Kod BHP i dobro diferentovanih PCa bila je veća ekspresija ER&beta; u bazalnim ćelijama nego u sekretornim. Lo&scaron;e diferentovani PCa prikazali su smanjenje ekspresije ER&beta; u bazalnim ćelijama. Ukupna ćelijska ekspresija ER&beta; predstavlja složen i ponekad moguće paradoksalan nalaz, na osnovu čega primarni PCa zadržava ekspresiju ovog receptora, ali ipak značajno nižu u poređenju sa benignim epitelom i premalignim lezijama. Ovaj nalaz podupire stanovi&scaron;te o antiproliferativnoj ulozi ER&beta; u tkivu prostate.</p> / <p>Adenocarcinoma of the prostate (PCa) is the most common cancer in men. High-grade prostatic intraepithelial neoplasia (HGPIN) are lesions that precede to invasive carcinomas and include complete absence of basal cells and stromal invasion by malignant acini. Estrogen receptor &szlig;(ER&szlig;) is located in the nuclei of basal and secretory cells and partly in stromal ones.The aim of the research is to describe and localize ER&szlig; in different morphological lesions: prostate hyperplasia (BPH), PIN and PCa with different Gleason score. It is assumed that pre-cancerous lesions in different stages of their evolution not correlate completely with the expression ER&szlig;. LGPIN shows expression, while there is no expression in HGPIN. It is also an assumption that the expression ER&szlig; exists in most medium differentiated PCa, and that the expression of this receptor loses with increasing of Gleason score. Five groups of patient were investigated: control group with BPH and four experimental groups (PIN and 3 different groups of PCa). The study was conducted on men of different ages in the period 2010-2012. None of the patients received prior hormonal therapy. Sextant prostate biopsy were stained on ER&szlig; (Novocastra). ER&szlig; expression is shown through the score: 0 = zero; 1 = &lt;1%; 2 = 1-10%; 3 = 11-33%; 4 = 34-66%; 5 =&gt; 66%. Positive fibroblasts and endothelial cells were used for comparison. Reduced expression was observed in malignant and premalignant lesions of the prostate versus BPH. ER&szlig; expression in the epithelial cells of acini was the weakest in well-differentiated PCa. In BPH and well differentiated PCa was greater expression in the basal cells than in secretory ones. Poorly differentiated PCa showed a decreased ER&szlig; expression in basal cells. Total cellular expression of ER&szlig; is a complex and sometimes paradoxical finding on the basis of which the primary PCa retains expression of this receptor, but significantly lower compared to benign epithelium and premalignant lesions. This finding supports the antiproliferative role of ER&szlig; in prostate tissue.</p>

INTRODUÇÃO: O câncer de próstata (CaP) é a neoplasia mais comum no homem depois do câncer de pele não-melanoma. É uma doença de comportamento heterogêneo e a hereditariedade é um dos principais fatores de risco. Recentes estudos do genoma humano revelaram numerosas regiões de susceptibilidade associada com a doença. Os Polimorfismos de Nucleotídeo Único (SNPs) são variantes de risco genéticos associados com uma série de doenças incluindo o câncer. Considerando que a história familiar constitui fator de risco para o desenvolvimento do CaP, acredita-se que a identificação de polimorfismos envolvidos nesse processo possa ter um papel relevante para auxiliar no desenvolvimento de ferramentas alternativas para a detecção precoce e para a definição do prognóstico desta neoplasia. OBJETIVOS: Análise dos polimorfismos genéticos relacionados ao desenvolvimento do câncer de próstata em indivíduos com história familiar para a doença. Além disso, correlacionar a frequência dos polimorfismos e os fatores prognósticos, tais como: nível do antígeno prostático específico (PSA), escore de Gleason, estadiamento patológico e recidiva bioquímica. MÉTODOS: Neste estudo foi analisado a frequência de 20 SNPs em 255 pacientes, sendo 185 pacientes portadores de câncer de próstata e 70 grupo controle. No grupo diagnosticado com CaP, tinham 97 casos esporádicos e 72 com histórico familiar (dois parentes de primeiro grau). O grupo controle foi composto por homens sem diagnóstico de CaP que estavam em prevenção com parâmetros como psa e toque retal normais. Todos foram submetidos a extração do DNA e o genótipo avaliado através da técnica de reação em cadeia da polimerase quantitativa em tempo real. A análise estatística foi realizada comparando a genotipagem e a frequência alélica entre os grupos, utilizando o teste de qui-quadrado com valor de significância menor ou igual a 0,05. RESULTADOS: Dois dos 20 polimorfismos apresentaram associação significativa com o CaP hereditário, o SNP rs7931342 (11q13.2) onde o genótipo homozigoto foi cinco vezes mais frequente (p=0,01) e o rs10090154 (8q24) onde o alelo polimórfico aumenta duas vezes e meia a chance de CaP hereditário (OR=2,67; p=0,04). O rs2660753 (3p12.1) mostrou relevância para o CaP esporádico onde a presença do alelo polimórfico tem um efeito protetor para o desenvolvimento da doença (OR=0,50; p=0,01). Com relação a associação dos fatores prognósticos, foram encontrados achados significativos para o SNP rs620861 (8q24) onde o genótipo homozigoto selvagem está associado com menor escore de Gleason (p=0,04). O polimorfismo rs1447295 (8q24) está relacionado a um melhor prognóstico, pois foi mais frequente nos pacientes com estadiamento localizado, sem recidiva bioquímica e com sobrevida livre de recidiva mais longa. CONCLUSÃO: Nosso estudo evidenciou dois SNP (rs7931342 e rs10090154) como fator de risco para o desenvolvimento de CaP hereditário. Com relação o CaP esporádico, a presença do alelo polimórfico do rs2660753 demonstra um efeito protetor. Por último, comparando os fatores prognósticos, tanto o genótipo homozigoto selvagem do rs620861 como o homozigoto polimórfico do rs1447295 conferem um bom prognóstico / INTRODUCTION: Prostate cancer (PCa) is the most common non-cutaneous malignancy among men. It is a heterogeneous disease and heredity is one of the strongest risk factors. Recent studies of the human genome revealed numerous susceptibility regions associated with the disease. Several single nucleotide polymorphisms (SNPs) have been implicated in the risk of developing the prostate cancer. Considering that the family history is a risk factor for PCa development, we believe that identification of polymorphisms involved in these processes may have an important role to help in the development of alternative tools for early detection and to define the prognosis of this neoplasm. OBJECTIVES: Analysis of genetic polymorphisms associated with the development of prostate cancer in patients with family history of prostate cancer. Moreover, correlate the frequency of polymorphisms and prognostic factors such as PSA level, Gleason score, pathological stage and biochemical recurrence. METHODS: In this study, the frequency of 20 genetic single nucleotide polymorphisms were analyzed in 255 patients, 185 patients with prostate cancer and 70 control. In the group diagnosed with PCa, they had 97 sporadic cases and 72 had a family history (two first-degree relatives). The control group consisted of men with no diagnosis of PCa who were on prevention with parameters such as normal psa and digital rectal exam. All were subjected to DNA extraction and genotype assessed by real-time polymerase chain reaction technique. Statistical analysis was performed by comparing the genotype and allele frequency between the groups using the chi-square test with significance or equal to 0.05. RESULTS: Two of the 20 polymorphisms were significantly associated with hereditary CaP, the rs7931342 (11q13.2) that the homozygote genotype was five times more common (p=0.01) and rs10090154 (8q24) that the polymorphic allele increase two times and a half the risk of hereditary PCa (OR = 0.5; p = 0.04). The rs2660753 (3p12.1) showed association with sporadic PCa where the presence of polymorphic allele has a protective effect on the PCa development (OR = 2,67, p = 0.01). Regarding the association of prognostic factors, we found significant findings to the SNP rs620861 (8q24) that the wild homozygote genotype is associated with lower Gleason score (p = 0.04). The rs1447295 polymorphism (8q24) is related to a better prognosis because it was more frequent in patients with localized stage, without biochemical recurrence and longer recurrence-free survival. CONCLUSIONS: Our study showed two SNPs (rs7931342 and rs10090154) as a risk factor for hereditary PCa development. Regarding the sporadic PCa, the polymorphic allele presence of rs2660753 demonstrated a protective effect. Finally, comparing the prognostic factors, both wild homozygote genotype of rs620861 and polymorphic homozygote of rs1447295 offer a good prognosis

INTRODUÇÃO - O câncer de próstata é a doença maligna mais diagnosticada no mundo, desconsiderando os tumores de pele. A prostatectomia radical tem sido considerada a opção terapêutica preferida no tratamento do câncer clinicamente localizado. O conhecimento da anatomia neurovascular da próstata é de extrema importância para a obtenção de resultados funcionais satisfatórios sem prejuízo aos fins oncológicos. A maioria dos estudos anatômicos já publicados utilizou próstatas removidas cirurgicamente - espécime de prostatectomia radical. Estudos realizados com próstata de cadáveres são importantes por permitir a ressecção em bloco dos órgãos pélvicos, possibilitando uma melhor avaliação da neuroanatomia periprostática. O objetivo deste estudo é avaliar detalhadamente a distribuição das fibras nervosas periprostáticas em cadáveres humanos. MÉTODOS - Foram obtidos 10 blocos viscerais de cadáveres com idade superior a 50 anos constituídos por bexiga, próstata e fáscias adjacentes, uretra proximal e reto. Após fixação em formalina, o bloco visceral foi fatiado transversalmente em fatias de aproximadamente 5 mm. Cada fatia foi dividida em quatro setores, tendo a uretra como eixo central e encaminhada para inclusão em parafina, secção com micrótomo padrão e confecção das lâminas coradas em HE. A cápsula prostática (CP) foi identificada e demarcada e áreas de avaliação foram definidas conforme distância da CP, designadas como distância D1(até 2,5 mm), D2 (2,5 a 5 mm) e D3(maior de 5 mm). Em cada região da próstata (região apical, região média e região basal) as fibras nervosas foram avaliadas de acordo com a distribuição circunferencial nos segmentos anterior, ântero-laterais direito e esquerdo, póstero-laterais direito e esquerdo e posterior e conforme a distância da CP - distâncias D1, D2 e D3. RESULTADOS - Foram identificados 29.275 fibras nervosas, das quais 3.274 (11,18%) foram detectados na região apical, 11.997 (40,98%) na região média e 14.004 (47,84%) na região basal da próstata. Na região basal, aproximadamente, 46% das fibras nervosas foram identificadas nas faces póstero-laterais, 20% na face posterior da próstata e menos de 9% na face anterior. Houve predomínio numérico de FN nas áreas mais distantes, com 46,62% delas sendo identificadas em D3. Na região média, 45% das fibras nervosas se distribuíram nos segmentos anteriores a ântero-laterais. Cerca de 45% se distribuíram igualmente nas faces póstero-laterais direita e esquerda e apenas 10% na face posterior. Nesta região, observou-se que mais de 60% das FN estavam dispostas nas áreas D1. Na região apical, também se observou predomínio de fibras nervosas nos segmentos que compõem as faces posteriores e póstero-laterais (61%). Aproximadamente, 24% das FN se distribuíram igualmente entre as faces ântero-laterais direita e esquerda e quase 15% estavam distribuídas na face anterior do ápice prostático. Mais da metade delas encontravam-se distribuídas a até 2,5 mm da cápsula prostática. Nos segmentos anteriores e ântero-laterais, o predomínio de FN nas áreas D1 foi mais acentuado do que nos outros segmentos. Na avaliação geral, a quantidade relativa de fibras nervosas identificadas nas áreas D1 variou de maneira inversamente proporcional à quantidade relativa de FN das áreas D3. Nas áreas demarcadas entre 2,5 mm e 5 mm da CP (D2), a quantidade relativa de fibras nervosas se manteve relativamente constante. CONCLUSÕES - No sentido craniocaudal, a quantidade de fibras nervosas diminui progressivamente, a distribuição circunferencial se modifica, com aumento da quantidade relativa de fibras nervosas nas faces anteriores e a distribuição em relação à distância da próstata também se modifica, com aumento da quantidade relativa de fibras nervosas nas áreas mais próximas da próstata. A quantidade relativa de fibras nervosas dispostas a uma distância mínima de 2,5 mm da cápsula prostática se mantém constante / INTRODUCTION - Prostate cancer is the most commonly diagnosed malignancy in the world, excluding skin tumors. The incidence of localized tumor increased substantially after the PSA era. Consequently, the percentage of men treated by radical surgery for clinically localized prostate cancer increased. The exact paths of periprostatic nerves have been under debate over the last decades. In the present study, the topographic distribution of nerves around the prostate and their relative distances from the prostatic capsule were analyzed in male cadaver visceral blocs. METHODS - The pelvic organs from ten fresh male cadavers were removed and serial sectioned en bloc for histological investigation. The prostatic capsule was identified, and distances of 2.5 mm and 5 mm from the prostate were demarked with lines. We quantified the number of nerve fibers present in each subsector of each slide and recorded their position relative to the prostatic capsule. RESULTS - A total of 29,275 nerve fibers were identified, of which 3,274 (11.18%) were detected in the apex, 11,997 (40.98%) in the middle and 14,004 (47.83%) in the basal area of the prostate. The majority of nerves were identified in the posterolateral and posterior surfaces of the prostate. At the apical region, the percentage of nerve fibers identified in the anterior region was higher. The nerves identified at the apex were mainly located up to 2.5 mm from the prostate. This proximity to the prostate was specifically observed in the ântero-lateral and anterior sectors. In the craniocaudal sense, the percentage of nerves identified between 2.5 and 5 mm from the prostatic capsule remained constant. CONCLUSIONS - A significant number of nerve fibers were present in the anterior and ântero-lateral positions, especially at the apex. The anterior nerves were closer to the prostate. This proximity suggests that the anterior nerves may participate in local physiology. It is likely that the safe distance of 2.5 mm from all surfaces of the prostate may be related to cavernous fiber preservation

by Lau Hoi Lun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 83-96). / Abstract also in Chinese. / Abstract --- p.i / Acknowledgements --- p.iv / Abbreviations --- p.v / Table of contents --- p.vi / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Endocrine control of normal and abnormal growth of prostate --- p.1 / Chapter 1.1.1 --- Androgen regulation of prostate gland --- p.1 / Chapter 1.1.2 --- Estrogen regulation of prostate gland --- p.4 / Chapter 1.2 --- Gonadotropin-releasing hormone plays a central role in reproduction --- p.6 / Chapter 1.2.1 --- GnRH gene --- p.7 / Chapter 1.2.2 --- GnRH receptor --- p.9 / Chapter 1.3 --- Therapeutic strategies using GnRH analogs to treat prostate cancer --- p.12 / Chapter 1.4 --- Expression of GnRH or its receptor in reproductive tissues --- p.12 / Chapter 1.4.1 --- Expression of GnRH in reproductive --- p.13 / Chapter 1.4.2 --- Expression of GnRH and its receptor in pituitary and reproductive tissues --- p.13 / Chapter 1.5 --- Animal models for the study of prostate cancer --- p.15 / Chapter 1.5.1 --- Nobel rat inducible model --- p.15 / Chapter 1.5.2 --- Androgen dependent rat Dunning prostatic tumor --- p.16 / Chapter 1.5.3 --- Androgen-independent prostatic carcinoma line of Noble rat --- p.18 / Chapter 1.6 --- Aim of study --- p.18 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Origin and supply of Nobel rat --- p.20 / Chapter 2.2 --- Induction of dysplasia in Nobel rat prostate gland by long-term treatment with steroids --- p.20 / Chapter 2.2.1 --- Chemicals --- p.20 / Chapter 2.2.2 --- Preparation of steroid hormone-filled Silastic tubings --- p.20 / Chapter 2.2.3 --- Surgical implantation of Silastic® tubings --- p.21 / Chapter 2.2.4 --- Protocols of hormonal treatments --- p.21 / Chapter 2.3 --- Androgen- dependent Dunning rat prostatic adenocarcinoma --- p.22 / Chapter 2.4 --- Androgen- independent prostatic carcinoma line (ALT) of Noble rat --- p.22 / Chapter 2.5 --- Detection of mRNA expression of gonadotropin- releasing hormone (GnRH) in normal and neoplastic rat prostates --- p.23 / Chapter 2.5.1 --- Preparation of tissue for total RNA extraction --- p.23 / Chapter 2.5.2 --- Total RNA extraction --- p.24 / Chapter 2.5.3 --- Reverse-transcription Polymerase Chain Reaction (RT-PCR) --- p.25 / Chapter 2.5.4 --- Purification of DNA fragments from agarose gels --- p.27 / Chapter 2.5.5 --- Subcloning of DNA into vector --- p.27 / Chapter 2.5.6 --- Nucleotide sequencing --- p.30 / Chapter 2.5.7 --- Southern blot analysis --- p.32 / Chapter 2.5.7.1 --- Southern blotting --- p.32 / Chapter 2.5.7.2 --- Preparation of α-32P-dCTP labelled GnRH probe --- p.32 / Chapter 2.5.7.3 --- Hybridization --- p.33 / Chapter 2.6 --- Detection of mRNA expression of gonadotropin-releasing hormone receptor (GnRH-R) in normal and neoplastic rat prostates --- p.34 / Chapter 2.6.1 --- Cloning of GnRH-R cDNA and synthesis of its probe --- p.34 / Chapter 2.6.2 --- Detection of GnRH receptor mRNA expression in normal and dysplastic Nobel rat prostates by Southern blot --- p.36 / Chapter 2.6.3 --- Detection of GnRH receptor mRNA expression in Dunning tumor --- p.37 / Chapter 2.6.4 --- Detection of the GnRH receptor mRNA expression in AIT tumor by RT-PCR --- p.37 / Chapter Chapter 3 --- Results / Chapter 3.1 --- Detection of mRNA expression of gonadotropin-releasing hormone (GnRH) in normal and neoplastic rat prostates --- p.38 / Chapter 3.1.1 --- Reverse -transcription Polymerase Chain Reaction (RT-PCR) --- p.38 / Chapter 3.1.2 --- Purification of DNA fragments amplified by PCR from the agarose gel --- p.38 / Chapter 3.1.3 --- Subcloning of DNA into vector --- p.39 / Chapter 3.1.4 --- Nucleotide sequencing --- p.39 / Chapter 3.1.5 --- Southern-blot analysis --- p.39 / Chapter 3.2 --- Detection of gonadotropin-releasing hormone receptor mRNA expression in normal and neoplastic rat prostates --- p.40 / Chapter 3.2.1 --- Cloning of gonadotropin-releasing hormone receptor (GnRH) cDNA and synthesis of probe from the normal Noble rat pituitary gland --- p.40 / Chapter 3.2.2 --- Detection of GnRH receptor mRNA expression in normal and dysplastic Nobel rat prostates --- p.42 / Chapter 3.2.3 --- Detection of GnRH receptor mRNA expression in rat Dunning tumor by PCR --- p.43 / Chapter 3.2.4 --- Detection of GnRH receptor mRNA expression in AIT tumor --- p.43 / Chapter Chapter 4 --- Discussion / Chapter 4.1 --- Detection of mRNA expression of gonadotropin-releasing releasing hormone(GnRH) in normal and neoplastic rat prostates --- p.69 / Chapter 4.1.1 --- Expression of GnRH mRNA in normal Nobel rat prostate gland --- p.69 / Chapter 4.1.2 --- Expression of GnRH mRNA in dysplastic Nobel rat prostate --- p.71 / Chapter 4.1.3 --- Expression of GnRH mRNA in androgen-dependent rat Dunning prostatic tumor --- p.72 / Chapter 4.1.4 --- Expression of GnRH mRNA in AIT tumor --- p.74 / Chapter 4.2 --- Detection of GnRH receptor in normal and dysplastic rat prostates --- p.75 / Chapter 4.2.1 --- Negative expression of GnRH receptor in normal and dysplastic Nobel in rat prostates --- p.75 / Chapter 4.2.2 --- Positive expression of GnRH receptor mRNA in rat Dunning tumor --- p.77 / Chapter 4.2.3 --- Negative expression of GnRH receptor mRNA in ALT tumor --- p.78 / Chapter Chapter 5 --- Summary and Conclusions --- p.80 / References --- p.83

Choi, Mei Yuk. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 79-89). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstract --- p.iii / Abstract (Chinese) --- p.v / List of Abbreviations --- p.vii / List of Figures --- p.viii / Chapter Chapter 1 --- Introduction / Chapter 1.1. --- General introduction of cancer --- p.1 / Chapter 1.2. --- Overview of apoptosis --- p.2 / Chapter 1.2.1. --- The extrinsic death receptor pathway --- p.4 / Chapter 1.2.2. --- The intrinsic mitochondrial pathway --- p.4 / Chapter 1.2.3. --- Cross-talk between the intrinsic and extrinsic pathways --- p.5 / Chapter 1.3. --- Overview of selenium --- p.6 / Chapter 1.3.1. --- Selenium and prostate cancer --- p.7 / Chapter i. --- Epidemiological studies --- p.7 / Chapter ii. --- Clinical trials --- p.8 / Chapter iii. --- Preclinical investigations --- p.10 / Chapter a. --- in vivo studies --- p.11 / Chapter b. --- in vitro studies --- p.12 / Chapter c. --- selenocystine and prostate cancer --- p.13 / Chapter 1.4. --- Objective --- p.15 / Chapter Chapter 2 --- Materials and methods / Chapter 2.1. --- Materials --- p.18 / Chapter 2.2. --- Methods --- p.19 / Chapter 2.2.1. --- Cell culture --- p.19 / Chapter 2.2.2. --- MTT assay --- p.19 / Chapter 2.2.3. --- Cell cycle distribution analysis --- p.20 / Chapter 2.2.4. --- TUNEL assay and DAPI staining --- p.20 / Chapter 2.2.5. --- Evaluation of mitochondrial membrane potential (ΔΨm) --- p.21 / Chapter 2.2.6. --- Measurement of superoxide generation (DHE assay) --- p.22 / Chapter 2.2.7. --- Inhibition of superoxide generation --- p.22 / Chapter 2.2.8. --- Western blot analysis --- p.23 / Chapter 2.2.9. --- Statistical analysis --- p.24 / Chapter Chapter 3 --- Results / Chapter 3.1. --- The antiproliferatvie effect of SeC on LNCaP and PC-3 cells --- p.25 / Chapter 3.2. --- The role of caspases in SeC-induced apoptosis --- p.34 / Chapter 3.3. --- The effect of SeC on the mitochondrial membrane potential --- p.39 / Chapter 3.4. --- The involvement of p53 in SeC-treated LNCaP cells --- p.44 / Chapter 3.5. --- MAPK and PI3K/Akt signaling pathways --- p.47 / Chapter 3.6. --- The role of superoxide in SeC-induced apoptosis --- p.52 / Chapter Chapter 4 --- Discussion --- p.62 / Chapter Chapter 5 --- Conclusion --- p.74 / References --- p.79

INTRODUÇÃO: O Câncer de Próstata (CaP) é a sexta neoplasia mais comum no mundo correspondendo a aproximadamente 10% do total de cânceres. No Brasil, o CaP é a neoplasia maligna não cutânea mais comum entre os homens. A hereditariedade é um dos principais fatores de risco do CaP, que se caracteriza pela herança de mutações em genes de susceptibilidade de alta penetrância que quando transmitidos aos descendentes aumentam o risco de desenvolvimento de tumores. Os andrógenos e estrógenos influenciam o desenvolvimento, maturação e manutenção da próstata, afetando a proliferação e a diferenciação. Isso tem despertado grande interesse no papel desses hormônios esteroides no desenvolvimento e manutenção tanto da próstata normal quanto maligna. Os Polimorfismos de Nucleotídeo Único (SNPs) são variantes de risco genéticos associados com uma série de doenças, incluindo o câncer. Considerando que a história familiar e que os componentes hormonais constituem fatores de risco para o desenvolvimento do CaP, acredita-se que a identificação de polimorfismos envolvidos nesses processos possa ter um papel relevante para auxiliar no desenvolvimento de ferramentas alternativas para a detecção precoce e para a definição do prognóstico desta neoplasia. OBJETIVOS: Analisar polimorfismos (SNPs) relacionados com histórico familiar e com fatores hormonais em amostras de sangue de pacientes com CaP e em homens saudáveis. Além disso, correlacionar os resultados da genotipagem com parâmetros clínico-patológicos. MÉTODOS: O estudo foi composto por 185 pacientes diagnosticados com CaP, sendo 97 casos esporádicos e 72 com histórico familiar (dois parentes de primeiro grau). O grupo controle foi composto por 70 amostras de sangue de indivíduos saudáveis, que comprovadamente não possuíam CaP e fazem acompanhamento com intuito preventivo. Foram selecionados 13 polimorfismos para análise: rs10486567, rs10993994, rs9364554, rs5945572, rs2735839, rs4430796, rs7501939, rs138213197, rs1271572, rs2987983, rs8072254, rs4919743 e rs3808330. A genotipagem foi realizada através da técnica de PCR em tempo real (qPCR) e correlacionada com o histórico familiar de CaP, PSA pré-operatório, graduação histológica de Gleason e estadiamento patológico. RESULTADOS: Analisamos a frequência dos polimorfismos selecionados e encontramos as seguintes correlações em nossos casos: os SNPs rs10486567 e rs9364554 aumentam a chance de desenvolvimento do CaP enquanto que o SNP rs8072254 diminui o risco. Com relação à hereditariedade, o SNP rs1271571 apresentou associação com o CaP esporádico. Na comparação com os fatores prognósticos encontramos que o SNP rs3808330 foi mais frequente em indivíduos que possuíam PSA < 10; o SNP rs7501939 foi mais frequente em indivíduos com menor escore de Gleason e ausência de recidiva e o SNP rs5945572 foi mais frequente em indivíduos com menor escore de Gleason na peça. CONCLUSÕES: De uma forma geral encontramos polimorfismos que parecem ter um papel relevante no desenvolvimento do CaP, na transmissão familiar e a fatores prognósticos. Estes importantes polimorfismos, ainda não haviam sido estudados na população brasileira e nosso trabalho identificou correlações ainda não demonstradas na literatura / BACKGROUND: Prostate Cancer (PCa) is the sixth most common cancer worldwide accounting for around 10% of all cancers. In Brazil, the PCa is the most common non-skin malignancy among men. Heredity is one of the main risk factors for PCa, which is characterized by mutations of heritage in highpenetrance susceptibility genes that when transmitted to offspring increase the risk of tumor development. Androgens and estrogens influence the development, maturation and maintenance of the prostate, affect proliferation and differentiation. This has aroused great interest in the role of these steroid hormones in the development and maintenance of both normal and malignant prostate. Single Nucleotide Polymorphisms (SNPs) are variants of genetic risk associated with a number of diseases including cancer. Considering that the family history and hormonal components are risk factors for the development of PCa, we believe that the identification of polymorphisms involved in these processes may have an important role to assist in the development of alternative tools for early detection and to define the prognosis of this cancer. OBJECTIVES: To analyze polymorphisms (SNPs) associated with family history and hormonal factors in patient blood samples with PCa and in healthy men. In addition, to correlate the results of genotyping with clinical-pathological parameters. METHODS: The study consisted of 185 patients diagnosed with PCa, divided into 97 sporadic cases and 72 with a family history. The control group consisted of 70 blood samples from healthy individuals who had no proven PCa and do it for preventive purposes. We selected 13 polymorphisms for analysis: rs10486567, rs10993994, rs9364554, rs5945572, rs2735839, rs4430796, rs7501939, rs138213197, rs1271572, rs2987983, rs8072254, rs4919743 and rs3808330. Genotyping was performed by PCR in real time (qRT -PCR) and correlated with family history of PCa, preoperative PSA, Gleason histologic grading and pathological staging. RESULTS: We analyzed the frequency of the selected polymorphisms and found the following correlations in our cases: SNPs rs10486567 and rs9364554 increase the chance of developing PCa while the SNP rs8072254 decreases the risk. Regarding heredity, the SNP rs1271571 presented association with sporadic PCa. In comparison with the prognostic factors we found that the SNP rs3808330 was more frequent in patients who had PSA < 10; SNP rs7501939 was more frequent in patients with lower Gleason score and no recurrence and the SNP rs5945572 was more frequent in subjects with lower Gleason score on the surgical specimen. CONCLUSIONS: In general we found that polymorphisms that appear to have a relevant role in the development of PCa in family transmission and in prognostic factors. These important polymorphisms had not yet been studied in the Brazilian population and our work has identified correlations yet not demonstrated in the literature

Serum PSA (Prostate Specific Antigen) level is used for prediction of prostatic carcinoma, but it suffers from weak sensitivity and specificity. We applied logistic regression, artificial neural networks, decision tree, and genetic algorithm to prostate cancer prediction problem to design a model for Turkish population. A hybrid model of logistic regression and decision tree has been designed. The model could prevent 33 biopsies (4.4% of our patients who have PSA level between 0 and 10) from our data set without a loss from sensitivity. The prepared online decision support tool and a questionnaire were published on a website. Fifty urologists have completed the questionnaire. Cronbach&rsquo / s alpha was 0.770. On a five graded Likert scale, the mean score of &ldquo / attitude to computer use in healthcare&rdquo / (ACH) was 4.2. The mean of eight responses related to the online tool (Attitude to Decision Support Tool / ADST), was 3.7. ADST was correlated with ACH (r=0.351, p=0.013). Physicians who have positive attitude to computer use in healthcare tend to use the tool (r=0.459, p=0.001). The first factor influencing the opinions of the urologists was the attitude of the user to computer use in healthcare, the other factor was the attitude of the user to the decision support tool itself. To increase the acceptance, education and training of physicians in the use of information technologies in healthcare, informing users about the logic of the decision support tool, and redesigning the system according to user feedback may be helpful.

Thesis (Ph. D.)--University of Oklahoma. / Bibliography: leaves 147-164.

Dissertation (Ph.D.)--University of South Florida, 2008. / Includes vita. Includes bibliographical references. Also available online.