by Yu Hon Ming. / Thesis (M.Sc.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 70-74). / Abstract --- p.ii / Acknowledgements --- p.v / Abbreviations --- p.vi / List of tables and figures --- p.viii / Chapter Chapter I. --- Introduction / Chapter 1. --- Background --- p.1 / Chapter 2. --- Physiology of prostate --- p.3 / Chapter 2.1. --- Prostate --- p.3 / Chapter 2.2. --- Embryonic development of prostate --- p.3 / Chapter 2.3. --- Anatomy of prostate --- p.5 / Chapter 2.4. --- The role of steroids in the growth of prostate --- p.7 / Chapter 2.4.1. --- T --- p.7 / Chapter 2.4.2. --- DHT --- p.8 / Chapter 2.5. --- 5α-reductase --- p.10 / Chapter 3. --- Pathophysiology of BPH --- p.11 / Chapter 3.1. --- Anatomic progression of BPH --- p.11 / Chapter 3.2. --- Epidemiology of BPH --- p.11 / Chapter 3.3. --- Pathogenesis of BPH --- p.12 / Chapter 3.4. --- Clinical manifestations of BPH --- p.13 / Chapter 3.5. --- Diagnosis of BPH --- p.14 / Chapter 4. --- Treatment of BPH --- p.15 / Chapter 4.1. --- a-adrenergic antagonist --- p.16 / Chapter 4.2. --- DHT hypothesis --- p.16 / Chapter 4.3. --- 5α-reductase inhibitor --- p.17 / Chapter 5. --- Radioimmunoassay of DHT --- p.21 / Chapter 6. --- Objectives --- p.22 / Chapter Chapter II. --- Materials and methods --- p.23 / Chapter 1. --- Materials and methods for development of specific RIA for serum DHT --- p.23 / Chapter 1.1. --- Materials --- p.23 / Chapter 1.2. --- Methods --- p.23 / Chapter 1.2.1. --- Antiserum for DHT-RIA --- p.23 / Chapter 1.2.1.1. --- Optimal antibody titre and dilution curve for DHT-RIA --- p.24 / Chapter 1.2.1.2. --- Cross-reactivity with related steroids --- p.25 / Chapter 1.2.2. --- KMnO4 treatment of T antiserum for DHT-RIA --- p.26 / Chapter 1.2.2.1. --- Optimization of KMnO4 treatment --- p.26 / Chapter 1.2.2.2. --- Upper limit of oxidizing power of 0.5% KMnO4 --- p.27 / Chapter 1.2.2.3. --- Efficiency and background effect of 0.5% KMnO4 oxidation on DHT-RIA --- p.28 / Chapter 1.2.3. --- Characteristic of DHT-RIA --- p.29 / Chapter 1.2.3.1. --- Standard preparation --- p.29 / Chapter 1.2.3.2. --- Blank preparation --- p.30 / Chapter 1.2.3.3. --- Control preparation --- p.31 / Chapter 1.2.3.4. --- Sample preparation --- p.31 / Chapter 1.2.3.5. --- Dextran-coated charcoal --- p.31 / Chapter 1.2.3.6. --- DHT-RIA procedure --- p.32 / Chapter 1.2.3.7. --- Sensitivity test for DHT-RIA --- p.33 / Chapter 1.2.3.8. --- Linearity test for DHT-RIA --- p.33 / Chapter 1.2.3.9. --- Recovery test for DHT-RIA --- p.34 / Chapter 1.2.3.10. --- Precision test for DHT-RIA --- p.34 / Chapter 2. --- Materials and methods for establishing reference range of DHT in Chinese males and females --- p.35 / Chapter 2.1. --- Samples --- p.35 / Chapter 2.2. --- Methods --- p.35 / Chapter 2.2.1. --- DHT determination --- p.35 / Chapter 2.2.2. --- T determination --- p.36 / Chapter 2.2.3. --- Cholesterol determination --- p.37 / Chapter 3. --- Materials and methods for a small clinical trial on the usefulness of 5α-reductase inhibitor (finasteride) in the treatment of BPH --- p.38 / Chapter 3.1. --- Samples --- p.38 / Chapter 3.2. --- Methods --- p.38 / Chapter 4. --- Statistical analysis --- p.39 / Chapter Chapter III. --- Result --- p.40 / Chapter 1. --- DHT-RIA --- p.40 / Chapter 1.1. --- Antiserum for DHT-RIA / Chapter 1.1.1. --- Optimal antibody titre for DHT-RIA --- p.40 / Chapter 1.1.2. --- Cross reactivity with related steroids --- p.42 / Chapter 1.2. --- KMnO4 treatment of T antiserum for DHT-RIA --- p.43 / Chapter 1.2.1. --- Optimization of KMnO4 treatment --- p.43 / Chapter 1.2.2. --- Upper limit of oxidizing power of 0.5% KMnO4 --- p.44 / Chapter 1.2.3. --- Efficiency and background effect of 0.5% KMnO4 treatment on DHT-RIA --- p.45 / Chapter 1.3. --- Characterization of the DHT-RIA --- p.47 / Chapter 2. --- Reference range for DHT in Chinese males and females --- p.51 / Chapter 2.1. --- Reference range of DHT in three different study groups --- p.51 / Chapter 2.2. --- Ratio of DHT/T in Chinese males and Caucasian males --- p.52 / Chapter 2.3. --- Correlation studies --- p.53 / Chapter 3. --- Results of small clinical trial --- p.54 / Chapter 3.1. --- Mean serum analyte changes during the course --- p.54 / Chapter 3.2. --- Changes in analyte concentrations during the course --- p.56 / Chapter 3.3. --- Difference in mean serum analytes between BPH males predose level and normal males with and without age matching --- p.57 / Chapter 3.4. --- "The response, percentage change of DHT and DHT/T ratio of individual patient during the course of study" --- p.58 / Chapter Chapter IV. --- Discussion --- p.59 / Chapter 1. --- Radioimmunoassay for DHT --- p.59 / Chapter 2 . --- Establishment of DHT reference range in Chinese males and females --- p.62 / Chapter 3. --- The usefulness of 5a reductase inhibitor in the treatment of BPH --- p.65 / Chapter Chapter V. --- Conclusion --- p.69 / References --- p.70

Les cancers du sein hormono-indépendants et HER2 non amplifiés, dites triples négatives (TN), représentent 30 % des tumeurs du sein. Elles sont de mauvais pronostic et ne bénéficient pas actuellement de thérapeutiques ciblées.Les tumeurs du sein apocrine moléculaire (AM) sont des tumeurs qui se caractérisent par l’absence de récepteurs hormonaux et la présence du récepteur aux androgènes (RA) dont la voie de signalisation est activée. Dans 50 % des cas elles sont HER2 négatives, et entrent alors dans la catégorie des TN.Le rôle de cette signalisation androgénique est mal connu. Cependant elle apparait comme une cible thérapeutique intéressante dans ce groupe de tumeur ne bénéficiant pas des thérapies ciblées. De plus peu de modèles précliniques représentatifs des tumeurs AM ont été publiés.Dans ce projet nous avons exploré le RA et sa signalisation à partir de modèles de lignées cellulaires.Nous avons d’abord caractérisé ces modèles sur le plan mutationnel en séquençant le RA et en recherchant des altérations moléculaires dans les voies de signalisation des récepteurs hormonaux et des facteurs de croissance puis sur le plan transcritpomique en vérifiant la présence d’une signature AM moléculaire et en recherchant des variants d’épissage du RA.Nous avons ensuite pu mettre en évidence une régulation de la prolifération et de la clonogénicité de ces lignées après blocage du RA et de sa voie de signalisation par invalidation de son expression par siRNA.Nous avons finalement pu vérifier dans plusieurs de ces modèles qu’un traitement par différents agonistes et antagonistes ciblant le RA permettait de moduler la prolifération et l’expression des gènes cibles du RA. / Hormone-independent and non-amplified HER2 breast cancers, or triple negative (TN), represent 30% of breast tumors. They have a poor prognosis and do not currently benefit from targeted therapies.Molecular apocrine (MA) breast cancers are characterized by the absence of hormone receptors and the presence of the androgen receptor (AR) which signaling pathway is activated. In 50% of cases they are HER2 negative, and then fall into the category of TN.The role of the androgen signaling is poorly understood. However the AR appears as an attractive therapeutic target in this group of tumors without targeted therapies. Moreover few preclinical models of MA have been published.In this project we explored the AR and its signaling in cell lines models.We first characterized these models on the mutational level by sequencing the AR and seeking molecular alterations in the signaling pathways of hormone receptors and growth factors. Then on the transcritpomique level by checking the presence of a molecular MA signature and seeking the AR splice variants.We then could highlight a regulation of cell proliferation and clonogenicity in these cell lines by blocking the AR and its signaling pathway with siRNAs.We were finally able to check in several of these models that treatment with different agonists and antagonists targeting the AR allowed to modulate cell proliferation and the expression of AR target genes.

Abstract Altogether 406 patients were included in five studies, and all patients were examined and controlled in the Oulu University Hospital during the years 1991–1998. Twenty-nine patients with mixed-type erectile dysfunction (ED) were recruited into a randomized, controlled, double-blind crossover comparison of placebo and high-dose yohimbine hydrochloride (36 mg per day orally). Positive clinical responses were obtained in 44% of the patients during yohimbine treatment and in 48% during placebo treatment. Thirty patients with ED underwent an intracavernosal injection test (ICI) using three different active agents (prostaglandin E1(PGE1), papaverine hydrochloride (PV), moxisylyte (MS)) and physiological saline. PGE1 produced significantly better rigidity than either PV or MS. Sixty-nine patients with ED who had started ICI therapy with PGE1 at least three years previously were invited to a control examination to find out the long-term outcome of this treatment and to evaluate the patients' overall satisfaction with their sexual life. 46.4% of the patients had discontinued PGE1 therapy, the mean time of using PGE1 having been 23.3 months (range 0–48 months). 34.8% of the patients reported that their own spontaneous erections had improved during the PGE1 therapy. The sexual functions of 155 patients with benign prostatic hyperplasia (BPH) were evaluated before TURP and 6 and 12 months afterwards with questionnaires. Only 26% of the patients had completely satisfactory erections before TURP, while 22% had satisfactory erections 6 months later and 24% 12 months later. The majority of patients (about 70%) were satisfied with their sexual life both before and after the procedure. 123 men with symptoms of andropause participated in a randomized, placebo-controlled study to assess the effects of dihydrotestosterone (DHT) gel in men with andropausal symptoms. The drug was administered transdermally once a day during six months. Early morning erections improved significantly (p < 0.003) in the DHT group by the three-month control, the ability to maintain erections was better, and there was also a positive effect on libido. In the patients with a elevated (> 12) international index of the prostatic symptoms score (I-PSS) before DHT treatment, I-PSS decreased from 17.7 to 12.3 points. As a conclusion yohimbine hydrochloride is no better than placebo in the treatment of patients with mixed-type ED. PGE1, PV and MS are well tolerated, and PGE1 was shown to be the most effective drug of the three. ICI therapy with PGE1 in long-term use is safe and effective. Sexual functions in men did not change after TURP, and this group of aging men were fairly satisfied with their sexual life despite of the fact that they had some ED and one third of the patients had not had intercourse during the previous year. Transdermal administration of DHT in aging men improves sexual function.

ABSTRACT Evidence from knockout studies in male mice and from experiments in male rats,in which expression of the estrogen receptor alpha (ERα) gene was inhibited in the medial preoptic area (MPO), suggests that ERα is important in the control of male rat mating behavior. Therefore, in this experiment, we tested the hypothesis that activation of ERα in the MPO is sufficient to maintain mating behavior in castrated male rats receiving subcutaneously (s.c.) dihydrotestosterone (DHT), a non-aromatizable androgen. Accordingly, castrated rats treated with DHT s.c. received MPO implants of either: (i) propyl-pyrazole-triol (PPT) (Stauffer, et al 2000; Katzenellenbogen, et al 2000), a selective ERα agonist, (ii) E2 (positive controls) or (iii) cholesterol (negative controls)and sexual behavior was monitored. PPT was as effective as E2 at maintaining mating behavior suggesting that, in the MPO, ERα is sufficient to mediate responses to E2 that underlie male rat mating behavior.

Abstract A hypothesis is created that an association between androgenetic alopecia (AGA) and serious cardiovascular events, such as myocardial infarction and fatal ischaemic heart disease has been reported, but the mechanism explaining this association has remained unclear. The aim of this study was to analyze the relationship between insulin resistance, (coronary) artery disease and AGA. Moreover, a hypothesis on the role of electromagnetic cell adhesion in the development of AGA is presented. In the present series of men aged 19–50 years (n = 154) with early (<  35 years of age), significant AGA of at least grade 3 (vertex) in the Hamilton classification modified by Norwood (Norwood 1975) was hyperinsulinaemia encountered twice as often as on age-matched controls. Other signs of the insulin resistance syndrome, such as obesity, lipid lowering and antihypertensive drugs were also found to correlate with early AGA. In a population-based case-control study, male patients living a small rural town who had undergone an urgent or elective coronary revascularization procedure (n =  85) and their age-matched controls were analysed after stratification by age at operation and hair status. The findings showed AGA to be more common coronary artery disease and early AGA as those with early coronary artery disease. In a population aged 63 years (n = 541, 217 men), neck circumference was found to correlate with the conventional anthropometric indicators of insulin resistance and with elevated serum insulin in both genders, which means that neck circumference is a simple anthropometric indicator of android type obesity and insulin resistance. In the same female population other factors of insulin resistance (whr, waist circumference, serum insulin level and microalbuminuria) were associated with marked (grade 2 or 3 on a modified Ludwig scale) hair loss. Paternal heredity was clearly characteristic of AGA in both genders, particularly of early AGA in men. We present a hypothesis that the overactive androgen state inhibits cell mitosis in the dermal papilla of the hair follicle and contributes to a weaker electromagnetic attraction between the undifferentiated germ cells and the dermal papilla and also to a shortened anagen phase of the hair growth cycle. Insulin resistance has an additional pathogenic role in the excessive miniaturization of the hair follicle. As a conclusion, along with android obesity, early alopecia can be considered a sign of insulin resistance and a possible risk factor for an early onset of coronary artery disease. Timely intervention in the risk factors may help to slow down or prevent the development of arterial disease and possibly also to alleviate the cosmetic and psychosocial consequences of hair loss. / Tiivistelmä Insuliiniresistenssin, (sepel)valtimotaudin ja AGA:n välillä on yhteyksiä. Taustalla olevat patomekanismit ovat kuitenkin epäselviä. Tässä väitöskirjatyössä tutkittiin insuliiniresistenssin ja (sepel)valtimotaudin suhdetta AGA:an. Lisäksi luotiin hypoteesi sähkömagneettisen soluadheesion roolista AGA:n kehittymisessä. Aineiston 19–50-vuotiailla miehillä (n = 154), joilla oli varhainen (< 35 v), merkittävä, vähintään kolmannen (vertex) asteen AGA Norwoodin modifioiman Hamiltonin luokituksen mukaan (Norwood 1975) seerumin insuliinipitoisuus oli suurentunut liki kaksi kertaa useammin kuin samanikäisillä verrokeilla. Myös muiden insuliiniresistenssioireyhtymään liitettyjen vaaratekijöiden, kuten ylipainon, havaittiin liittyvän varhaiseen AGA:an. Pienen maaseutukaupungin kaikki sepelvaltimoiden revaskularisaatioon joutuneet miehet (n = 85) analysoitiin toimenpiteeseen joutumisiän ja hiusstatuksen mukaan. Tulokset osoittavat AGA:n olevan yhteydessä sepelvaltimotautiin ja varhaisen AGA:n varhaiseen sepelvaltimotautiin. Aineiston 63-vuotiailla (n = 541, miehiä 217) kaulan ympärysmitan todettiin korreloivan selvästi antropometrisiin, insuliiniresistenssiä kuvaaviin mittoihin ja seerumin insuliinipitoisuuden kasvuun sekä miehillä että naisilla. Kaulan ympärysmitta soveltuu siten käytettäväksi antropometrisena mittana androidityyppisen ylipainon ja insuliiniresistenssin selvittämisessä. Saman väestöotoksen naisilla tehdyssä tutkimuksessa havaittiin muiden insuliiniresistenssin osatekijöiden (vyötärö-lantiosuhteen, vyötärön ympärysmitan, seerumin insuliinipitoisuuden ja mikroalbuminurian) liittyvän huomattavaan hiustenlähtöön (asteet II ja III modifioidulla Ludwigin skaalalla). AGA:ssa isän suvun perimän vaikutus oli selvä molemmilla sukupuolilla. Se oli voimakas erityisesti miesten varhaisessa AGA:ssa. Laatimamme hypoteesin mukaan suuri androgeenipitoisuus estää dermaalipapillan solujen mitoosia ja heikentää sähkömagneettista vetovoimaa. Tällöin hiusfollikkelin solujen määrää vähenee ja hiuksen kasvuvaihe lyhenee haittaavasti. Insuliiniresistenssillä on hypoteesin mukaan toissijainen rooli hiusfollikkelin pienenemisprosessissa. Aikaista hiustenlähtöä androidin ylipainon ohella voidaan pitää insuliiniresistenssin merkkinä ja riskinä sepelvaltimotaudin tavanomaista aiempaan ilmaantumiseen. Puuttumalla ajoissa vaaratekijöihin valtimotaudin kehittymistä voidaan hidastaa tai estää ja ehkä myös vähentää kosmeettisesti ja psykososiaalisesti haittaavaa hiusten menetystä.

Introdução: A presença de tecido prostático em pacientes 46,XX portadoras de hiperplasia congênita das supra-renais (HCSR) já foi relatada por alguns autores. Acredita-se que o desenvolvimento deste tecido decorre do estímulo androgênico, através da dihidrotestosterona, sobre as glândulas parauretrais de Skene destas pacientes. Estas glândulas, presentes em todas as meninas, possuem homologia histológica e enzimática com a próstata masculina. Além da presença de tecido prostático nestas pacientes, houve dois relatos de alterações neste tecido, sendo o primeiro o de uma paciente 46,XX portadora de deficiência de 21-hidroxilase, que desenvolveu uma hiperplasia benigna prostática e outra com a mesma deficiência enzimática que aos 62 anos apresentou adenocarcinoma de próstata. Objetivos: 1. Verificar a ocorrência de tecido prostático nas pacientes acima de 6 anos, portadoras da forma clássica de hiperplasia congênita das supra-renais, com cariótipo 46,XX, em acompanhamento no ambulatório de endocrinologia pediátrica do Instituto da Criança; 2. Analisar a sensibilidade e especificidade do antígeno prostático específico (PSA) e da dihidrotestosterona das pacientes com HCSR em relação à detecção do tecido prostático através da ressonância nuclear magnética da região pélvica; 3. Correlacionar os níveis de PSA com os níveis de testosterona e dihidrotestosterona das pacientes portadoras de HCSR. Casuística: Constituiu-se de 32 pacientes 46,XX portadoras da forma clássica de hiperplasia congênita das suprarenais (31 com deficiência de 21-hidroxilase e uma com deficiência de 11- hidroxilase), com uma média de idade de 11,8 +/- 4,2 anos e um grupo controle que incluiu 10 meninas e 10 meninos sem hiperplasia congênita das supra-renais, com média de idade de 9,8 +/- 1,9 anos e 9,3 +/- 2,7 anos respectivamente. Os pacientes do grupo controle faziam acompanhamento no ambulatório de endocrinologia pediátrica por baixa estatura ou puberdade precoce. Métodos: O padrão-ouro utilizado para a detecção de tecido prostático foi a ressonância magnética de região pélvica, que foi realizada em todos os pacientes. Também foram dosados o antígeno prostático específico, a 17-hidroxiprogesterona, o 21-desoxicortisol, o 11- desoxicortisol, a androstenediona, a testosterona e a dihidrotestosterona. Resultados: A detecção do tecido prostático nas ressonâncias de região pélvica foi obtida em 5 pacientes com hiperplasia congênita das supra-renais. O antígeno prostático específico mostrou uma sensibilidade de 100% e especificidade de 88,9% para um ponto de corte de 0,1 ng/mL. A dihidrotestosterona revelou uma sensibilidade de 100% e especificidade de 60% para um ponto de corte de 27,7 ng/dL. Houve uma correlação positiva entre os níveis de PSA com os níveis de T e entre os níveis de PSA e DHT. Conclusões: A ocorrência de tecido prostático nas pacientes 46,XX portadoras de hiperplasia congênita das supra-renais estudadas foi de 15,6%. O PSA mostrou ser um valioso marcador de tecido prostático nas pacientes com HCSR. De acordo com os achados, a pesquisa de tecido prostático deve ser considerada em todas as pacientes portadoras da forma clássica de HCSR / Introduction: The presence of prostatic tissue in 46,XX congenital adrenal hyperplasia (CAH) patients has already been reported. The development of this tissue is due to androgenic stimulation in Skenes paraurethral glands by dihydrotestosterone. These glands have histological and enzymatic homology with the prostate. So far, two cases with alterations of prostatic tissues have been reported: one 46,XX patient with 21-hydroxylase deficiency developed benign prostatic hyperplasia and another with the same enzymatic deficiency, at age of 62 years, had adenocarcinoma of prostate. Objectives: 1.To describe the presence of prostatic tissue in 46, XX patients with the classical form of congenital adrenal hyperplasia 2. To evaluate the sensitivity and specificity of prostatic specific antigen (PSA) and dihydrotestosterone with regard to the detection of prostatic tissue in pelvic MRI. 3. To correlate prostate specific antigen levels with testosterone and dihydrotestosterone levels in girls with congenital adrenal hyperplasia. Patients and Methods: Among our CAH patients followed at our Unit, we selected 32 children and adolescents 46,XX, from 6 to 22 years of age, with the classical form of congenital adrenal hyperplasia (mean age 11.8 +/- 4.2); 31 patients had 21- hydroxylase deficiency and one 11-hydroxylase deficiency. Control group included 10 boys (mean age 9.3 +/- 2.7) and 10 girls (mean age 9.8 +/- 1.9) without CAH. Pelvic MRI (taken as the “gold standard method”) was performed in all patients to detect prostatic tissue. Prostate specific antigen, 17-hydroxyprogesterone, 21- deoxycortisol, 11-deoxycortisol, androstenedione, testosterone and dihydrotestosterone were measured in all patients. Results: Five congenital adrenal hyperplasia girls showed image of prostatic tissue on pelvic MRI. Prostate specific antigen had sensitivity and specificity of 100.0% and 88.9%, respectively, taking 0.1 ng/mL as the cutoff level. The dihydrotestosterone had a sensitivity and specificity of 100% and 60%, respectively, with a cutoff level of 27.7 ng/dL. There was a positive correlation between PSA levels and testosterone and dihydrotestosterone levels. Conclusions: The incidence of prostatic tissue in 46,XX patients with the classical form of congenital adrenal hyperplasia was 15.6%. PSA demonstrated to be a good marker of prostatic tissue in these patients. Based on this study, we have to be cautious when treating a girl with the classical form of CAH and the search for prostatic tissue must be considered in every patient

Introdução: A presença de tecido prostático em pacientes 46,XX portadoras de hiperplasia congênita das supra-renais (HCSR) já foi relatada por alguns autores. Acredita-se que o desenvolvimento deste tecido decorre do estímulo androgênico, através da dihidrotestosterona, sobre as glândulas parauretrais de Skene destas pacientes. Estas glândulas, presentes em todas as meninas, possuem homologia histológica e enzimática com a próstata masculina. Além da presença de tecido prostático nestas pacientes, houve dois relatos de alterações neste tecido, sendo o primeiro o de uma paciente 46,XX portadora de deficiência de 21-hidroxilase, que desenvolveu uma hiperplasia benigna prostática e outra com a mesma deficiência enzimática que aos 62 anos apresentou adenocarcinoma de próstata. Objetivos: 1. Verificar a ocorrência de tecido prostático nas pacientes acima de 6 anos, portadoras da forma clássica de hiperplasia congênita das supra-renais, com cariótipo 46,XX, em acompanhamento no ambulatório de endocrinologia pediátrica do Instituto da Criança; 2. Analisar a sensibilidade e especificidade do antígeno prostático específico (PSA) e da dihidrotestosterona das pacientes com HCSR em relação à detecção do tecido prostático através da ressonância nuclear magnética da região pélvica; 3. Correlacionar os níveis de PSA com os níveis de testosterona e dihidrotestosterona das pacientes portadoras de HCSR. Casuística: Constituiu-se de 32 pacientes 46,XX portadoras da forma clássica de hiperplasia congênita das suprarenais (31 com deficiência de 21-hidroxilase e uma com deficiência de 11- hidroxilase), com uma média de idade de 11,8 +/- 4,2 anos e um grupo controle que incluiu 10 meninas e 10 meninos sem hiperplasia congênita das supra-renais, com média de idade de 9,8 +/- 1,9 anos e 9,3 +/- 2,7 anos respectivamente. Os pacientes do grupo controle faziam acompanhamento no ambulatório de endocrinologia pediátrica por baixa estatura ou puberdade precoce. Métodos: O padrão-ouro utilizado para a detecção de tecido prostático foi a ressonância magnética de região pélvica, que foi realizada em todos os pacientes. Também foram dosados o antígeno prostático específico, a 17-hidroxiprogesterona, o 21-desoxicortisol, o 11- desoxicortisol, a androstenediona, a testosterona e a dihidrotestosterona. Resultados: A detecção do tecido prostático nas ressonâncias de região pélvica foi obtida em 5 pacientes com hiperplasia congênita das supra-renais. O antígeno prostático específico mostrou uma sensibilidade de 100% e especificidade de 88,9% para um ponto de corte de 0,1 ng/mL. A dihidrotestosterona revelou uma sensibilidade de 100% e especificidade de 60% para um ponto de corte de 27,7 ng/dL. Houve uma correlação positiva entre os níveis de PSA com os níveis de T e entre os níveis de PSA e DHT. Conclusões: A ocorrência de tecido prostático nas pacientes 46,XX portadoras de hiperplasia congênita das supra-renais estudadas foi de 15,6%. O PSA mostrou ser um valioso marcador de tecido prostático nas pacientes com HCSR. De acordo com os achados, a pesquisa de tecido prostático deve ser considerada em todas as pacientes portadoras da forma clássica de HCSR / Introduction: The presence of prostatic tissue in 46,XX congenital adrenal hyperplasia (CAH) patients has already been reported. The development of this tissue is due to androgenic stimulation in Skenes paraurethral glands by dihydrotestosterone. These glands have histological and enzymatic homology with the prostate. So far, two cases with alterations of prostatic tissues have been reported: one 46,XX patient with 21-hydroxylase deficiency developed benign prostatic hyperplasia and another with the same enzymatic deficiency, at age of 62 years, had adenocarcinoma of prostate. Objectives: 1.To describe the presence of prostatic tissue in 46, XX patients with the classical form of congenital adrenal hyperplasia 2. To evaluate the sensitivity and specificity of prostatic specific antigen (PSA) and dihydrotestosterone with regard to the detection of prostatic tissue in pelvic MRI. 3. To correlate prostate specific antigen levels with testosterone and dihydrotestosterone levels in girls with congenital adrenal hyperplasia. Patients and Methods: Among our CAH patients followed at our Unit, we selected 32 children and adolescents 46,XX, from 6 to 22 years of age, with the classical form of congenital adrenal hyperplasia (mean age 11.8 +/- 4.2); 31 patients had 21- hydroxylase deficiency and one 11-hydroxylase deficiency. Control group included 10 boys (mean age 9.3 +/- 2.7) and 10 girls (mean age 9.8 +/- 1.9) without CAH. Pelvic MRI (taken as the “gold standard method") was performed in all patients to detect prostatic tissue. Prostate specific antigen, 17-hydroxyprogesterone, 21- deoxycortisol, 11-deoxycortisol, androstenedione, testosterone and dihydrotestosterone were measured in all patients. Results: Five congenital adrenal hyperplasia girls showed image of prostatic tissue on pelvic MRI. Prostate specific antigen had sensitivity and specificity of 100.0% and 88.9%, respectively, taking 0.1 ng/mL as the cutoff level. The dihydrotestosterone had a sensitivity and specificity of 100% and 60%, respectively, with a cutoff level of 27.7 ng/dL. There was a positive correlation between PSA levels and testosterone and dihydrotestosterone levels. Conclusions: The incidence of prostatic tissue in 46,XX patients with the classical form of congenital adrenal hyperplasia was 15.6%. PSA demonstrated to be a good marker of prostatic tissue in these patients. Based on this study, we have to be cautious when treating a girl with the classical form of CAH and the search for prostatic tissue must be considered in every patient

No description available.

Prostate cancer (PCa) represents the most frequently diagnosed cancer and the second leading cause of cancer death in males. Initial development and progression of the disease is mainly regulated by androgens. However, the pathology of the disease may progress to a loss of hormone dependence, resulting in rapid growth and a metastatic phenotype. Invasion and metastasis of tumour cells results from the degradation of the basement membrane (BM) and extracellular matrix (ECM). The degradation of the BM and ECM is in part mediated by a family of proteinases called the matrix metalloproteinases (MMPs). Currently more than 20 members of the MMP family have been identified and they are further divided in to sub-classes according to their protein structure. Collectively, MMPs are capable of degrading essentially all ECM components. High expression of some MMPs correlates with a malignant phenotype of various tumours. This study focused on the expression and regulation of a sub-class of MMPs called the membrane-type MMPs (MT-MMPs) in PCa. To date 6 MT-MMPs have been identified and they are characterized by a transmembrane domain, followed by a short cytoplasmic tail (MT1-, MT2-, MT3- and MT5-MMPs) or a glycosylphosphatidylinositol (GPI) moiety (MT4- and MT6-MMPs). MT-MMPs are thought to play a key role in tumour cell invasion by virtue of their ability to activate MMP-2 (a secreted MMP, which is implicated in many metastatic tumours) and their direct degradation activity on ECM components. Elevated MT-MMP expression has been shown in breast, colon, skin, stomach, lung, pancreas and brain cancers. Until very recently there had been no studies conducted on MT-MMPs in PCa. The few studies preceding or occurring in parallel with this one, have mainly reported the mRNA expression of these enzymes in PCa. Most studies have focused on MT1-MMP. Thus, at the commencement of this project there were many unexplored aspects of the expression and regulation of the broader MT-MMP family in PCa. The aims of this study were to examine: 1 a) The expression of MT-MMPs in prostate cancer cell lines using RT-PCR and western blot analysis and b) expression of MT1-MMP and MT5-MMP in BPH (benign prostatic hyperplasia) and PCa clinical tissue sections by immunohistochemistry. 2) The regulation of MT1-MMP, MT3-MMP and MT5-MMP in PCa cell lines by Concanavalin A (Con A), phorbol-12-myristate 13-acetate (PMA), dihydrotestosterone (DHT) and insulin-like growth factors I and II (IGF I and IGF II) using western blot analysis. In this study RWPE1, a transformed but non-tumorigenic prostate cell line was used as a "normal" prostate cell model, ALVA-41 and LNCaP as androgen-dependent PCa cell models and DU-145 and PC-3 as androgen-independent PCa cell models. The mRNA expression for the 6 MT-MMPs was determined by RT-PCR. The results indicate that MT1- and MT3-MMP were detected in all cell lines. This is the first study to report MT1-MMP mRNA expression in LNCaP cells and MT3-MMP mRNA in DU-145 cells. MT2-MMP mRNA was detected in only LNCaP and DU-145 cells, whilst MT5-MMP was detected in PC-3, DU-145 and LNCaP cells. nterestingly, MT2-, MT4-, MT5- or MT6-MMP mRNA expression was not detected in the "normal" cell line RWPE1, perhaps indicating an induction in gene transcription in tumour cells. MT4-MMP mRNA was only detected in the androgen-independent cell lines, indicating a potential role in the invasion and metastasis processes of the aggressive androgen-independent PCa. In this study, very low expression of MT6-MMP was detected only in LNCaP and DU-145 cells. Previously there had been no reports on the expression of MT6-MMP in the normal or cancerous prostate. Due to the mRNA of MT1-, MT3- and MT5-MMPs being the predominant MT-MMPs expressed in the current study, and the availability of suitable antibodies against them, the protein expression of these three MT-MMPs was studied by western blot analysis. MT1-, MT3- and MT5-MMP protein expression was detected in the cell lysates and conditioned medium (CM) of RWPE1, LNCaP and PC-3 cells. For each MT-MMP, various protein species were detected including putative proforms, mature (active) forms, processed or fragmented forms as well as soluble or shed forms. The presence of soluble or shed forms of MT-MMPs in the CM of cultures of "normal" and PCa cells could imply one of the following mechanisms: ectodomain shedding by either extracellular sheddases, the secretion of intracellular processed proteins without the transmembrane domain, the release of membrane vesicles containing membrane-bound enzymes, or the presence of alternatively spliced mRNA, which gives rise to MT-MMPs without a transmembrane domain. Further characterization of these various forms, including their amino acid sequence, is required to fully elucidate their structural composition. Despite the detection of the mRNA, we did not detect the cell-associated proteins of MT1-MMP and MT5-MMP and only very low expression of MT3-MMP in DU-145 cells (CM of DU-145 cells were not screened for soluble forms of the enzymes). This is the first study to report MT5-MMP expression at the protein level in prostate derived cell lines. Immunohistochemistry was carried out on benign prostatic hyperplasia (BPH) and PCa clinical tissues using MT1- and MT5-MMP antibodies to determine their cellular localisation in benign and cancer glands. MT1- and MT5-MMPs were expressed in BPH and moderate and high grade PCa. MT1-MMP expression was highest in moderate grade cancer compared to BPH and high grade cancer. MT1-MMP expression was predominantly observed in the cytoplasm of secretory epithelial cells of both benign and cancer glands, although in cancer glands, some nuclear staining was also observed. Stromal expression of MT1-MMP was only observed in high grade cancer. This study is the first to report the immunolocalization of MT5-MMP outside the brain and in kidneys of diabetic patients. MT5-MMP was predominantly expressed in the cytoplasm of the secretory cells in benign glands. In the cancer glands, staining was heterogeneous with low to intense staining, mainly in the nuclei, plasma membrane and cytoplasm of secretory epithelial cells. Stromal expression of MT5-MMP was only observed in cancer tissues, particularly in high grade cancer. To study the regulation of MT-MMPs in PCa, we treated LNCaP and PC-3 cells, with either Con A, PMA, DHT or IGF-I and -II and studied the protein expression of MT1-, MT3- and MT5-MMPs by western blot analysis. Con A and PMA have been shown to stimulate MMP expression in other cell systems. Con A treatment showed a general increase in the protein expression of MT1-, MT3- and MT5-MMPs. By far the greatest induction by Con A observed was the nearly 4 fold increase in MT5-MMP expression caused by 40μg/mL Con A treatment of PC-3 cells. PMA treatment of LNCaP and PC-3 cells appeared to increase shedding or secretion of all three MT-MMPs in to the CM. This increase in the soluble forms corresponded to a decrease in cell-associated forms in LNCaP cells. Treatment of LNCaP with DHT alone and treatment of LNCaP and PC-3 cells with IGF-I and -II alone failed to detect any change in expression of MT1-MMP. The information gathered in this study on MT-MMPs with respect to cellular localization, expression levels and regulation by growth factors or chemicals that mimic their actions, will aid in our understanding of the role of MT-MMPs in PCa. This study provides strong preliminary data for further research, particularly with respect to functional studies of MT-MMPs in PCa. Understanding the processes which govern the actions of such proteins as these will provide potential insights into development of new management and therapeutic regimens to prevent cancer progression.