Ο καρκίνος του προστάτη αποτελεί σημαντικό πρόβλημα της δημόσιας υγείας ιδιαίτερα στο δυτικό κόσμο. Είναι ο πιο συχνός σπλαχνικός καρκίνος και ο δεύτερος πιο θανατηφόρος μετά τον καρκίνο του πνεύμονα. Η εισαγωγή και χρήση του PSA από τα τέλη της δεκαετίας του ’80 προσέθεσε ένα ισχυρό διαγνωστικό εργαλείο στα χέρια των κλινικών, το οποίο αναχαίτισε τη μέχρι τότε αυξητική τάση του αριθμού των θανάτων οφειλομένων στον καρκίνο του προστάτη. Το PSA ωστόσο χαρακτηρίζεται από χαμηλή ειδικότητα αλλά και ευαισθησία, χαρακτηριστικά που επιβάλλουν περιορισμούς στη χρήση του. Εξάλλου δεν προβλέπει το βαθμό της επιθετικότητας ενός αδενοκαρκινώματος του προστάτη, συμβάλοντας στο φαινόμενο της υπερδιάγνωσης αλλά κυρίως της υπερθεραπείας του καρκίνου του προστάτη. Κατά συνέπεια είναι απαραίτητη η ανάπτυξη μοριακών εργαλείων (δεικτών) οι οποίοι θα διαγιγνώσκουν τη νόσο με μεγαλύτερη ασφάλεια αλλά κυρίως θα μας δίνουν προγνωστικές πληροφορίες για τη βιολογική συμπεριφορά του όγκου, προλαμβάνοντας μια άσκοπη θεραπευτική παρέμβαση. Ενδιαφέρον πεδίο αναζήτησης τέτοιων δεικτών αποτελεί η χρωμοσωμική περιοχή 8q24. Ο μικροδορυφόρος DG8S737 εδράζεται στην περιοχή 8q24 και έχει δειχθεί, σε διαφορετικούς πληθυσμούς, ότι συγκεκριμένο αλληλόμορφό του σχετίζεται με επιθετικό, κλινικά σημαντικό καρκίνο του προστάτη. Στη συγκεκριμένη μελέτη ανιχνεύονται οι συχνότητας των αλληλομόρφων του μικροδορυφόρου DG8S737 σε μια ομάδα γενικού πληθυσμού (control) και σε μια ομάδα ασθενών οι οποίοι πάσχουν από επιθετικό καρκίνο του προστάτη. Τα αποτελέσματα επιβεβαιώνουν την τάση του αλληλομόρφου -8 να ανιχνεύεται συχνότερα σε ασθενείς παρά σε υγιείς. Εξάλλου παρατηρήθηκε για πρώτη φορά η ίδια τάση και για το αλληλόμορφο -10. Αυτά τα αποτελέσματα ενισχύουν το δυναμικό χρήσης του DG8S737 ως δείκτη για το μέτρο της επιθετικότητας του καρκίνου του προστάτη. / Prostate cancer represents a major public health issue in western world. It is the most frequently diagnosed visceral cancer whereas it is second in mortality. The use of PSA since the late 80s restrained the rising tendency of mortality of prostate cancer. PSA though, lacks in specificity. Besides it contributes to the phenomenon of overdiagnosis which leads to overtreatment of prostate cancer. Consequently it is necessary for novel biomarkers to emerge in order to diagnose more accurately and predict the aggressiveness of prostate cancer. The 8q24 region of chromosome 8 is a region which could harbor potential biomarkers for prostate cancer. The microsatellite DG8S737 in that region has a number of alleles, one of which has the tendency to be more often detected in patients with aggressive prostate cancer. We have studied the frequencies of alleles of DG8S737 in a group of patients with aggressive prostate cancer as well as in a group of control volunteers. The results confirm the findings of previous studies. The allele -8 of DG8S737 has been detected more often in patients than in healthy volunteers. A new finding is that allele -10 also is more frequently detected in the patients group rather than the control group. The results confirm previous findings and enforce the potential use of DG8S737 as novel biomarker for the aggressive prostate cancer.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23081号 / 医博第4708号 / 新制||医||1049(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 中本 裕士, 教授 小川 誠司, 教授 武田 俊一 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

1 ABSTRACT Introduction and aim of the study Circulating tumor cells (CTCs) are a promising tool of identifying patients with castration- resistant prostate cancer (CRPC) who will benefit from often demanding cytotoxic therapy. The aim of this work was to evaluate the prognostic significance of CTC in docetaxel-treated CRPC patients. During the project, we also tested the various methods of CTC cultivation and studied their genetic profile as well as the genetic profile of histological specimen at the time of diagnosis. Patients and methods A total of 39 patients who met the CRPC criteria and were indicated for docetaxel chemotherapy were included in the prospective study. Blood collection for CTC analysis was done in all patients before chemotherapy and on the first day of the fourth or fifth cycle of docetaxel. In parallel, CTCs were cultivated. Isolation and detection of CTC was done using the AdnaTest system, which consists of immunomagnetic separation and subsequent detection of mRNA from the CTC lysate. The primary objective of the study was to evaluate the overall survival (OS) of patients. Survival analysis was performed using the Kaplan-Meier method of estimating the survival distribution function. The impact of individual factors was tested using the Log-rank test, the Wilcoxon test and the Cox...

L'hormonothérapie (HTX) est un traitement couramment administré chez les hommes atteints d'un cancer de la prostate et qui peut avoir des répercussions sur leur bien-être émotionnel, en incluant une occurrence possible de dépression. Les études qui ont évalué la relation entre l'HTX et le développement de la dépression chez ces patients ont tantôt démontré une telle association et tantôt non. Une explication avancée serait que le cancer de la prostate et ses traitements puissent mener à des altérations dans les niveaux de testosterone et de Cortisol pouvant être associées à la dépression. Cependant, ces études comportaient plusieurs limites méthodologiques dont l'utilisation d'un devis de recherche transversal et l'absence d'un groupe contrôle approprié. La présente thèse compte deux articles empiriques issus de la même étude longitudinale menée auprès de 60 participants traités par radiothérapie seule (n = 32) ou combinée à de l'HTX (n = 28) évalués avant le début des traitements et à sept autres reprises sur une période de 16 mois. Cette thèse visait à répondre aux objectifs principaux suivants : (1) vérifier si l'introduction de l'HTX (et le retrait) de l'HTX étaient associés à une augmentation (et une diminution) des troubles dépressifs et de la sévérité de la dépression; (2) documenter l'évolution des niveaux sanguins de testosterone et de Cortisol dans chacun des groupes; et (3) évaluer la relation entre les niveaux hormonaux de testosterone et de Cortisol et la dépression. Globalement, les résultats obtenus suggèrent que l'HTX a eu un impact mineur sur le développement et la résolution de la dépression chez ces hommes. Par ailleurs, les niveaux de testosterone et de Cortisol ont diminué de façon significative dans les deux groupes de traitement. Néanmoins, la baisse de testosterone dans le groupe qui recevait de l'HTX n'était pas associée significativement à la dépression alors que des scores plus élevés de Cortisol étaient significativement reliés à des niveaux plus importants de dépression, et ce, dans les deux groupes de traitement. Ces résultats contribuent au développement d'une meilleure compréhension de la relation entre les traitements oncologiques, les changements hormonaux et la dépression. Il apparaît maintenant important d'investiguer davantage les différents mécanismes hormonaux et psychologiques sous-jacents au développement de la dépression chez cette population.

Thèse ou mémoire avec insertion d'articles / Contexte: Aucune étude n'a décrit la qualité de vie (QdV) des hommes à risque de cancer de la prostate (CaP) au Canada. Notre objectif visait à faire une analyse descriptive complète de la QdV dans une cohorte canadienne d'hommes à haut risque de développer un CaP, et à évaluer l'impact des problèmes urinaires et érectiles sur la QdV générale. Méthodes: La QdV a été récoltée chez 2053 hommes à risque de CaP participant à une étude observationnelle prospective multicentrique appelée BIOmarqueurs, Cancer de la Prostate, Prévention et Environnement (BIOCaPPE). Les participants ont rempli plusieurs questionnaires validés afin d'évaluer leur QdV générale(Échelle d'anxiété et de dépression [HADS] et Inventaire de 36 items forme courte [SF-36]), et celle spécifique au CaP (Score international des symptômes de la prostate [IPSS] et Inventaire de la santé sexuelle pour hommes [SHIM]). Résultats: Parmi tous les participants, 122 (6,1%) représentent des cas définitifs d'anxiété et 40 (2,0%) des cas définitifs de dépression; 1068 (53,9%) ont des symptômes urinaires modérés à sévères et 154 (7,8%) ont des symptômes de dysfonction érectile sévères. Les participants ont une QdV similaire à celle des hommes de la population générale, à l'exception de la fonction urinaire. Plus les symptômes urinaires et de dysfonction érectile sont sévères, moins bonne est la QdV générale. Les symptômes urinaires ont un plus grand impact sur la QdV générale que les symptômes de dysfonction érectile. Conclusions: Nos résultats suggèrent que la majorité des participants perçoivent leur QdV comme étant satisfaisante, bien que la majorité présentent des symptômes urinaires modérés à sévères. Les symptômes anxieux, dépressifs et érectiles sont moins fréquents. Les symptômes urinaires et de dysfonction érectile ont un impact négatif sur la QdV générale. Il s'agit de la première analyse de la QdV d'une cohorte canadienne d'hommes à risque de CaP / Background: No study has described the quality of life (QoL) of men at risk for prostate cancer (PCa) in Canada. Our objective was to conduct a comprehensive descriptive analysis of QoL in a Canadian cohort of men at risk of developing PCa, and to assess the impact of urinary and erectile symptoms on general QoL. Methods: QoL was collected in 2053 men at risk for PCa participating in a prospective multicenter observational study called BIOmarkers, Prostate Cancer, Prevention and Environment (BIOCaPPE). Participants completed several validated questionnaires to assess their general QoL (Anxiety and Depression Scale [HADS] and 36-item Short Form Survey [SF-36]), and PCa-specific QoL (International Prostate Symptom Score [IPSS] and Sexual Health Inventory for Men [SHIM]). Results: Of all participants, 122 (6.1%) are definite cases of anxiety and 40 (2.0%) definite cases of depression; 1068 (53.9%) have moderate to severe urinary symptoms and 154 (7.8%) have severe erectile dysfunction symptoms. Participants have a QoL similar to that of men in the general population, except for urinary function. More severe the urinary and erectile dysfunction symptoms are, worse the general QoL is. Urinary symptoms have a larger impact on general QoL than erectile dysfunction symptoms. Conclusions: Our results suggest the majority of participants perceived their QoL as satisfactory, although the majority have moderate to severe urinary symptoms. Anxiety, depressive, and erectile symptoms are less common. Urinary and erectile dysfunction symptoms have a negative impact on general QoL. This is the first analysis of the QoL of a Canadian cohort of men at risk of 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

Two areas of particular importance in prostate cancer progression are primary tumour development and metastasis. These processes involve a number of physiological events, the mediators of which are still being discovered and characterised. Serine proteases have been shown to play a major role in cancer invasion and metastasis. The recently discovered phenomenon of their activation of a receptor family known as the protease activated receptors (PARs) has extended their physiological role to that of signaling molecule. Several serine proteases are expressed by malignant prostate cancer cells, including members of the kallikreinrelated peptidase (KLK) serine protease family, and increasingly these are being shown to be associated with prostate cancer progression. KLK4 is highly expressed in the prostate and expression levels increase during prostate cancer progression. Critically, recent studies have implicated KLK4 in processes associated with cancer. For example, the ectopic over-expression of KLK4 in prostate cancer cell lines results in an increased ability of these cells to form colonies, proliferate and migrate. In addition, it has been demonstrated that KLK4 is a potential mediator of cellular interactions between prostate cancer cells and osteoblasts (bone forming cells). The ability of KLK4 to influence cellular behaviour is believed to be through the selective cleavage of specific substrates. Identification of relevant in vivo substrates of KLK4 is critical to understanding the pathophysiological roles of this enzyme. Significantly, recent reports have demonstrated that several members of the KLK family are able to activate PARs. The PARs are relatively new members of the seven transmembrane domain containing G protein coupled receptor (GPCR) family. PARs are activated through proteolytic cleavage of their N-terminus by serine proteases, the resulting nascent N-terminal binds intramolecularly to initiate receptor activation. PARs are involved in a number of patho-physiological processes, including vascular repair and inflammation, and a growing body of evidence suggests roles in cancer. While expression of PAR family members has been documented in several types of cancers, including prostate, the role of these GPCRs in prostate cancer development and progression is yet to be examined. Interestingly, several studies have suggested potential roles in cellular invasion through the induction of cytoskeletal reorganisation and expression of basement membrane-degrading enzymes. Accordingly, this program of research focussed on the activation of the PARs by the prostate cancer associated enzyme KLK4, cellular processing of activated PARs and the expression pattern of receptor and agonist in prostate cancer. For these studies KLK4 was purified from the conditioned media of stably transfected Sf9 insect cells expressing a construct containing the complete human KLK4 coding sequence in frame with a V5 epitope and poly-histidine encoding sequences. The first aspect of this study was the further characterisation of this recombinant zymogen form of KLK4. The recombinant KLK4 zymogen was demonstrated to be activatable by the metalloendopeptidase thermolysin and amino terminal sequencing indicated that thermolysin activated KLK4 had the predicted N-terminus of mature active KLK4 (31IINED). Critically, removal of the pro-region successfully generated a catalytically active enzyme, with comparable activity to a previously published recombinant KLK4 produced from S2 insect cells. The second aspect of this study was the activation of the PARs by KLK4 and the initiation of signal transduction. This study demonstrated that KLK4 can activate PAR-1 and PAR-2 to mobilise intracellular Ca2+, but failed to activate PAR-4. Further, KLK4 activated PAR-1 and PAR-2 over distinct concentration ranges, with KLK4 activation and mobilisation of Ca2+ demonstrating higher efficacy through PAR-2. Thus, the remainder of this study focussed on PAR-2. KLK4 was demonstrated to directly cleave a synthetic peptide that mimicked the PAR-2 Nterminal activation sequence. Further, KLK4 mediated Ca2+ mobilisation through PAR-2 was accompanied by the initiation of the extra-cellular regulated kinase (ERK) cascade. The specificity of intracellular signaling mediated through PAR-2 by KLK4 activation was demonstrated by siRNA mediated protein depletion, with a reduction in PAR-2 protein levels correlating to a reduction in KLK4 mediated Ca2+mobilisation and ERK phosphorylation. The third aspect of this study examined cellular processing of KLK4 activated PAR- 2 in a prostate cancer cell line. PAR-2 was demonstrated to be expressed by five prostate derived cell lines including the prostate cancer cell line PC-3. It was also demonstrated by flow cytometry and confocal microscopy analyses that activation of PC-3 cell surface PAR-2 by KLK4 leads to internalisation of this receptor in a time dependent manner. Critically, in vivo relevance of the interaction between KLK4 and PAR-2 was established by the observation of the co-expression of receptor and agonist in primary prostate cancer and prostate cancer bone lesion samples by immunohistochemical analysis. Based on the results of this study a number of exciting future studies have been proposed, including, delineating differences in KLK4 cellular signaling via PAR-1 and PAR-2 and the role of PAR-1 and PAR-2 activation by KLK4 in prostate cancer cells and bone cells in prostate cancer progression.

[Truncated abstract] In recent years a better understanding of the biology of PPAR , a nuclear transcription factor, has emerged, leading to a resurgence in targeting PPAR for chemotherapy. The family of synthetic PPAR agonists, the thiazolidinediones (TZDs), and non-steroidal anti-inflammatory drugs (NSAIDs) have been implicated in the inhibition of cell proliferation, apoptosis and cell cycle arrest of androgen-sensitive (LNCaP) and androgen-independent (PC-3 and DU145) prostate cancer cells generating much interest in their use for potential curative cancer therapies. In light of the potential use of TZDs and NSAIDs in prostate cancer prevention and their ability to induce inhibitory effects in vitro and in vivo, the first aim of this project was to undertake a comprehensive study of the effects of ciglitazone (TZD) and indomethacin (NSAID) on the androgen-independent prostate cancer cell line DU145, using standardised concentrations and time-points to compare the effects of TZDs and NSAIDs on cell proliferation, cell cycle and apoptosis. Treating the cells with either 10 µM ciglitazone or 10 µM indomethacin resulted in a time-dependent decrease in DU145 cell proliferation. The anti-proliferative effects were found to be in-part attributed to the slowing of cell progression through the G1/S-phase checkpoint of the cell cycle, and in the case of ciglitazone, apoptosis also played a role in its anti-proliferative effects in this cell line. Interestingly, although indomethacin failed to induce apoptosis, its antiproliferative effects were more potent than ciglitazone. The second aim of this project was to further investigate the underlying mechanisms responsible for the anti-proliferative effects of ciglitazone and indomethacin by evaluating their ability to modulate PPAR mRNA and protein expression, and to induce PPAR transcriptional activity. ... In addition, ligandinduced regulation of secreted frizzled related protein 4 (sFRP4) expression, a Wnt/ - catenin antagonists, was investigated. It was demonstrated that both ciglitazone and indomethacin attenuated Wnt/ -catenin signalling via the down-regulation of total - catenin levels within the cells, inhibition or slowing of the translocation of cytoplasmic -catenin into the nucleus and inhibition of cyclin–D1 expression An inverse relationship between PPAR and -catenin protein levels was also detected, suggesting that PPAR may directly bind to -catenin itself. sFRP4 expression was transiently upregulated by ciglitazone and indomethacin-treatment, suggesting that the antiproliferative effects of the ligands may be mediated in part through regulation of sFRP4 mRNA and protein levels. In summary, the anti-proliferative effects of ciglitazone and indomethacin on the androgen-independent prostate cancer cell line, DU145, described in this thesis are progressive steps in characterising the role of PPAR in prostate cancer cell proliferation. The identification of indomethacin as a more potent PPAR agonist than ciglitazone represents a novel target for the development of preventative strategies for advanced disease, and the relationship between PPAR and the Wnt/ -catenin signalling pathway provide an insight into the mechanisms involved in the anti-proliferative effects of ciglitazone and indomethacin. Further studies into this relationship would advance help identify novel preventative and curative therapeutic strategies for advanced prostate cancer.

Prostate cancer is one of the leading causes of death in men aged 40-55. Genistein isoflavone (4', 5', 7-trihydroxyisoflavone) is a dietary phytochemical with demonstrated anti-tumor activities in a variety of cancers. Topotecan Hydrochloride (Hycamtin) is an FDA-approved chemotherapy drug, primarily used for secondary treatment of ovarian,cervical and small cell lung cancers. This study was to demonstrate the potential anticancer activities and synergy of topotecan-genistein combination in LNCaP prostate cancer cells. The potential efficacy and mechanism of topotecan/genistein-induced cell death was investigated... Results: The overall data indicated that i) both genistein and topotecan induce cellular death in LNCaP cells, ii) topotecan-genistein combination was significantly more efficacious in reducing LNCaP cell viabiligy compared to either genistein or topotecan alone, iii) in all cases, cell death was primarily through apoptosis, via the activation of the intrinsic pathway, iv) ROS levels were increased and VEGF expression was diminished significantly with the topotecan-genistein combination treatment, v) genetic analysis of topotecan-genistein treatment groups showed changes in genetic expression levels in pathway specific apoptotic genes.... Conclusion: Treatments involving topotecan-genistein combination may prove to be an attractive alternative phytotherapy of adjuvant therapy for prostate cancer. / by Vanessa P. Hèormann. / Thesis (Ph.D.)--Florida Atlantic University, 2012. / Includes bibliography. / Mode of access: World Wide Web. / System requirements: Adobe Reader.

Indiana University-Purdue University Indianapolis (IUPUI) / This dissertation studies the effects of fatty acids on prostate cancer. Prostate cancer is one of the most common malignant diseases in males in the U.S. Because of the slow progression of this disease, early intervention methods, especially, dietary fatty acid interventions are considered very important to control the disease in early stages. This study describes how the depletion of the enzyme for endogenous fatty acid synthesis, fatty acid synthase, influences the expression of enzymes that metabolize dietary fatty acids and show how dietary fatty acids affect prostate cancer protein expression and function. Fatty acid synthase is an oncoprotein overexpressed in prostate cancer and its expression is suppressed with omega-3 fatty acid treatment. This study finds that the depletion of fatty acid synthase by siRNA knockdown induces suppression of cyclooxygenase-2 and fatty acid desaturase-1. Our results also show that fish oil (omega-3 fatty acid), but not oleic acid (omega-9 fatty acid), suppresses prostate cancer cell viability. Assessment of fatty acid synthesis activity indicates that oleic acid is a more potent inhibitor than fish oil of de novo fatty acid biosynthesis. In addition, the inhibition of its activity occurs over several days while its effects on cell viability occur within 24 hours. To better understand this relationship, label free LC-MS/MS based mass spectrometry was carried out to determine global proteomic and phosphoproteomic profiles of the prostate cell line PC3, with longitudinal treatment with fish oil or oleic acid. With short-term fish oil treatment, sequestosome-1was elevated. Prolonged treatment induced downregulation of microseminoprotein, a proinflammation factor, as well as proteins in the glycolysis pathway. In the phosphoproteomics study, we confidently identified 828 phosphopeptides from 361 phosphoproteins. Quantitative comparison between fish oil or oleic acid treated groups and the untreated group suggests that the fish oil induces changes in phosphorylation of proteins involved in the pathways associated with cell viability and metabolic processes, with fish oil inducing significant decreases in the levels of phospho-PDHA1Ser232 and phospho-PDHA1Ser300 and they were accompanied by an increase in PDH activity, suggesting a role for n-3 polyunsaturated fatty acids in controlling the balance between lipid and glucose oxidation.