As the most frequently diagnosed cancer in North American men, prostate cancer can progress to the androgen independent stage after initial response to androgen ablation therapy. The molecular mechanisms involved in the hormonal progression of prostate cancer are not completely understood. Here, we analyze changes in the transcriptome of prostate cancer cells at different stages of progression to reveal potential mechanisms. Applying Affymetrix GeneChip technology, we identified the transcriptomes in response to stimulation of androgen and PKA pathways in human prostate cancer cells. In addition to PSA, other common target genes were identified. Genes differentially expressed in response to androgen and stimulation of the PKA pathway in vitro were also differentially expressed during hormonal progression in vivo. Upon androgen stimulation, androgen receptor binds to a functional androgen response element within the promoter region of SESN1, a p53 targeted gene, and represses its expression. The expression of SESN1 was induced by castration in LNCaP xenografts, but the expression was eventually suppressed again in the androgen independent stage of prostate cancer. Knockdown of SESN1 promoted the proliferation of prostate cancer cells. Expression patterns of androgen-regulated genes in androgen independent tumours were revealed to be more similar to that from before castration than to the tumors under androgen ablation. The β-catenin, a potent coactivator of the androgen receptor, and Wnt pathway was deregulated in androgen-independent tumours. There was increased nuclear colocalization and interaction of androgen receptor and β-catenin with hormonal progression of prostate cancer. This study provides insight into hormonal effects on prostate cancer and possible pathways involved in the development of androgen independent disease, as well as potential therapeutic targets.

The mechanism by which prostate cancer progresses from androgen dependence (AD) to androgen independence/castration resistance (AI/CR) is currently a major focus of prostate cancer-related research. Prostate cancers that progress to a state of AI/CR are typically resistant to most standard types of treatments. Due to its primary role in driving normal prostate cell growth and proliferation, the androgen receptor (AR) is believed to play a key role in progression. Coregulators, or any proteins which may either enhance or abrogate AR activity, are considered to be one of the potential mechanisms by which AR function may become impaired. Cyclin G-associated kinase (GAK) was initially identified as a potential coregulator of AR in a Tup 1 repressed transactivation system. A LNCaP cDNA library was screened for proteins which interacted with the NH2-terminus of AR. GAK was isolated from three independent library clones using two different AR baits (AR 1-549 and AR 1-646). This interaction was confirmed via GST pulldown and coimmunoprecipitation experiments, and preliminary luciferase assays suggested that GAK activates AR in a hormone dependent manner. In this study, my objectives were to validate GAK’s role as a coregulator of AR and to determine if overexpressing GAK affects progression to AI. In vitro luciferase assays whereby GAK was either overexpressed or knocked down in both LNCaP and PC3 cells did not significantly affect AR activity. Xenograft experiments utilizing a doxycycline (DOX) inducible lentiviral LNCaP-GAK overexpressing stable cell line demonstrated that while GAK may not play a significant role in modulating AR activity, it may adopt a more subtle role enhancing tumour take and tumour volume growth rate in vivo. While these results could not confirm GAK to be a direct coregulator of AR, it is entirely possible that GAK may influence prostate cancer progression, albeit indirectly. Recent publications report a growing amount of evidence suggesting GAK’s involvement in the critical cellular process of clathrin coated vesicle endocytosis, the dysregulation of which could potentially indirectly affect AR regulated genes.

Les métalloprotéinases matricielles (MMPs) jouent un rôle crucial dans le développement malin des tumeurs. Elles dégradent la matrice extracellulaire (MEC) permettant ainsi la migration des cellules cancéreuses vers d'autres foyers. Ce processus d'invasion tumorale est appelé "processus métastasique". La MMP-9, appelée aussi gélatinase B, est fortement exprimée lors de ce processus. Son expression peut être régulée à différents niveaux via des voies de signalisation intracellulaires, notamment au niveau de l'expression du gène, mais aussi lors de la transcription, la stabilité de son ARNm, la traduction ou la sécrétion de la protéine. Des agents anti-tumoraux sont en cours de développement ciblant les différentes étapes du développement cancéreux. Le PCK3145, un peptide synthétique, dérivé de la PSP94, dirigé contre le cancer de la prostate avancé et métastasé est au stade d'essais cliniques en phase II. Ce peptide réduit chez la souris immunodéficiente la croissance des xénogreffes de tumeurs prostatiques humaines en diminuant les concentrations plasmatiques de la MMP-9. Nos travaux visent à élucider in vitro les mécanismes moléculaires impliqués dans la régulation des fonctions de la MMP-9 via l'effet anti-métastasique du PCK3145. Dans un premier temps, nous avons étudié l'interaction de la MMP-9 avec la surface de la cellule cancéreuse. Nous démontrons que le PCK3145 inhibe la sécrétion de la MMP-9 et déclenche le processus de clivage de CD44 de la surface cellulaire via une cascade de signalisation intracellulaire impliquant la GTPase RhoA. Dans un second lieu, nous avons étudié le mécanisme d'action du PCK3145 menant à l'inhibition de la sécrétion de la MMP-9. Nous démontrons que cette inhibition requiert le récepteur de la laminine à 67 kDa et est dépendante de l'activation de la voie des MAPKinases. Le PCK3145 entraine la diminution de la sécrétion de la MMP-9 via HuR, une protéine intracytoplasmique qui stabilise l'ARNm de la MMP-9 en se liant à sa séquence riche en AU. Les propriétés anti-métastasiques du PCK3145 peuvent être dirigées contre d'autres types de tumeurs, autres que le cancer de la prostate, liés à l'augmentation de la MMP-9 et à la dégradation de la MEC lors d'un processus métastasique. Par ailleurs, l'identification du mode d'action du PCK3145 via le récepteur de la laminine à 67 kDa permet de cibler des cancers dont le taux de ce récepteur est particulièrement élevé telle que la leucémie. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Cancer de la prostate, Métastase, Migration, Adhésion, MMP-9, HuR, ERK1/2, RhoA, CD44.

The early detection of prostate cancer plays a significant role in the success of treatment and outcome. To detect prostate cancer, imaging modalities such as TransRectal UltraSound (TRUS) and Magnetic Resonance Imaging (MRI) are relied on. MRI images are more comprehensible than TRUS images which are corrupted by noise such as speckles and shadowing. However, MRI screening is costly, often unavailable in many community hospitals, time consuming, and requires more patient preparation time. Therefore, TRUS is more popular for screening and biopsy guidance for prostate cancer. For these reasons, TRUS images are chosen in this research. Radiologists first segment the prostate image from ultrasound image and then identify the hypoechoic regions which are more likely to exhibit cancer and should be considered for biopsy. In this thesis, the focus is on prostate segmentation and on Regions of Interest (ROI)segmentation. First, the extraneous tissues surrounding the prostate gland are eliminated. Consequently, the process of detecting the cancerous regions is focused on the prostate gland only. Thus, the diagnosing process is significantly shortened. Also, segmentation techniques such as thresholding, region growing, classification, clustering, Markov random field models, artificial neural networks (ANNs), atlas-guided, and deformable models are investigated. In this dissertation, the deformable model technique is selected because it is capable of segmenting difficult images such as ultrasound images. Deformable models are classified as either parametric or geometric deformable models. For the prostate segmentation, one of the parametric deformable models, Gradient Vector Flow (GVF) deformable contour, is adopted because it is capable of segmenting the prostate gland, even if the initial contour is not close to the prostate boundary. The manual segmentation of ultrasound images not only consumes much time and effort, but also leads to operator-dependent results. Therefore, a fully automatic prostate segmentation algorithm is proposed based on knowledge-based rules. The new algorithm results are evaluated with respect to their manual outlining by using distance-based and area-based metrics. Also, the novel technique is compared with two well-known semi-automatic algorithms to illustrate its superiority. With hypothesis testing, the proposed algorithm is statistically superior to the other two algorithms. The newly developed algorithm is operator-independent and capable of accurately segmenting a prostate gland with any shape and orientation from the ultrasound image. The focus of the second part of the research is to locate the regions which are more prone to cancer. Although the parametric dynamic contour technique can readily segment a single region, it is not conducive for segmenting multiple regions, as required in the regions of interest (ROI) segmentation part. Since the number of regions is not known beforehand, the problem is stated as 3D one by using level set approach to handle the topology changes such as splitting and merging the contours. For the proposed ROI segmentation algorithm, one of the geometric deformable models, active contours without edges, is used. This technique is capable of segmenting the regions with either weak edges, or even, no edges at all. The results of the proposed ROI segmentation algorithm are compared with those of the two experts' manual marking. The results are also compared with the common regions manually marked by both experts and with the total regions marked by either expert. The proposed ROI segmentation algorithm is also evaluated by using region-based and pixel-based strategies. The evaluation results indicate that the proposed algorithm produces similar results to those of the experts' manual markings, but with the added advantages of being fast and reliable. This novel algorithm also detects some regions that have been missed by one expert but confirmed by the other. In conclusion, the two newly devised algorithms can assist experts in segmenting the prostate image and detecting the suspicious abnormal regions that should be considered for biopsy. This leads to the reduction the number of biopsies, early detection of the diseased regions, proper management, and possible reduction of death related to prostate cancer.

LIV-1, a zinc transporter, is an effector molecule downstream from soluble growth factors. This protein has been shown to promote epithelial-to-mesenchymal transition (EMT) in human pancreatic, breast, and prostate cancer cells. Despite the implication of LIV-1 in cancer growth and metastasis, there has been no study to determine the role of LIV-1 in prostate cancer progression. Moreover, there is no clear delineation of the molecular mechanism underlying LIV-1 function in cancer cells. In this study, we found increased LIV-1 expression in a progresssive manner in benign, PIN, primary and bone metastatic human prostate cancer. We characterized the mechanism by which LIV-1 drives prostate cancer EMT in an androgen-refractory human prostate cancer cell (ARCaP) bone metastasis model. LIV-1, when overexpressed in ARCaPE cells (derivative cells of ARCaP with epithelial phenotype), promoted EMT irreversibly. LIV-1 overexpressed ARCaPE cells had elevated levels of HB-EGF and matrix metalloproteinase (MMP) 2 and MMP 9 proteolytic enzyme activities, without affecting intracellular zinc concentration. The activation of MMPs resulted in the shedding of heparin binding-epidermal growth factor (HB-EGF) from ARCaPE cells, eliciting constitutive epidermal growth factor receptor (EGFR) phosphorylation and its downstream extracellular signal regulated kinase (ERK) signaling. Further investigation of the HB-EGF promoter revealed that both Stat3 and AP-1 controlled HB-EGF promoter activity. Ectopic LIV-1 overexpression induced AP-1 and Stat3 activation. Blockade of both Stat3 and AP-1 by specific inhibitors or dominant negative expression vectors diminished the HB-EGF promoter activity induced by LIV-1 overexpression. These results suggest that LIV-1 is involved in prostate cancer progression as an intracellular target of growth factor receptor signaling which promotes EMT and cancer metastasis. LIV-1 could be an attractive therapeutic target for the eradication of pre-existing human prostate cancer and bone and soft tissue metastases.

Sexuality is a sensitive topic in health care and is often interpreted through a natural scientific lens as just corresponding to sexual dysfunction and fertility problems. The purpose of this thesis was to describe sexuality and its outcomes in two cancer populations. Women with breast cancer and men with prostate cancer in all stages were invited to participate. In this thesis, these two populations are restricted to age groups between 45 and 65 years, since there are reasons to believe that younger people are more vulnerable to sexuality changes. Lifeworld, gender, and sexuality are three concepts of importance in this thesis and they are used from the viewpoint of nursing care. Phenomenological interviews (I, III) and focus group interviews (II, IV) were carried out with a total number of 46 informants. The EPP-method (Empirical Phenomenological Psychological) was used (I, III) in order to grasp the lived experience, and qualitative content analysis was used to analyse the seven focus groups (II, IV). The lifeworld experiences of those women and men were comparable. The changes brought by the cancer and its treatment were a threat to their very existence, their existential base of knowledge had gone and alienation occurred (I, III). For the women, this was illustrated through the metaphor of a bird which is pinioned and unable to fly anymore. For the men it was expressed in the essential meaning “to lose the elixir of life”. Both women and men suffered, sexuality changed from one day to another and they handled it individually. Changed body appearance, and feeling old and unattractive were, for the women, the dominating features, whilst for the men changed desire and erection problems were their main concerns. The findings from the group discussions (II, IV) elucidate the gendered differences in these two contexts. The aim of the women was to look healthy and attractive and for the men the ability to have an erection was important. Neither of these two groups of people was able to meet their aims. On the other hand, being diagnosed with a life-threatening disease they were not in a position to claim preserved sexuality. This opens up existential questions that need to be confirmed in health care. To succeed in this, a change of perspective is required in health care. It should be possible to use human science to the same extent as natural science in health care.

The early detection of prostate cancer plays a significant role in the success of treatment and outcome. To detect prostate cancer, imaging modalities such as TransRectal UltraSound (TRUS) and Magnetic Resonance Imaging (MRI) are relied on. MRI images are more comprehensible than TRUS images which are corrupted by noise such as speckles and shadowing. However, MRI screening is costly, often unavailable in many community hospitals, time consuming, and requires more patient preparation time. Therefore, TRUS is more popular for screening and biopsy guidance for prostate cancer. For these reasons, TRUS images are chosen in this research. Radiologists first segment the prostate image from ultrasound image and then identify the hypoechoic regions which are more likely to exhibit cancer and should be considered for biopsy. In this thesis, the focus is on prostate segmentation and on Regions of Interest (ROI)segmentation. First, the extraneous tissues surrounding the prostate gland are eliminated. Consequently, the process of detecting the cancerous regions is focused on the prostate gland only. Thus, the diagnosing process is significantly shortened. Also, segmentation techniques such as thresholding, region growing, classification, clustering, Markov random field models, artificial neural networks (ANNs), atlas-guided, and deformable models are investigated. In this dissertation, the deformable model technique is selected because it is capable of segmenting difficult images such as ultrasound images. Deformable models are classified as either parametric or geometric deformable models. For the prostate segmentation, one of the parametric deformable models, Gradient Vector Flow (GVF) deformable contour, is adopted because it is capable of segmenting the prostate gland, even if the initial contour is not close to the prostate boundary. The manual segmentation of ultrasound images not only consumes much time and effort, but also leads to operator-dependent results. Therefore, a fully automatic prostate segmentation algorithm is proposed based on knowledge-based rules. The new algorithm results are evaluated with respect to their manual outlining by using distance-based and area-based metrics. Also, the novel technique is compared with two well-known semi-automatic algorithms to illustrate its superiority. With hypothesis testing, the proposed algorithm is statistically superior to the other two algorithms. The newly developed algorithm is operator-independent and capable of accurately segmenting a prostate gland with any shape and orientation from the ultrasound image. The focus of the second part of the research is to locate the regions which are more prone to cancer. Although the parametric dynamic contour technique can readily segment a single region, it is not conducive for segmenting multiple regions, as required in the regions of interest (ROI) segmentation part. Since the number of regions is not known beforehand, the problem is stated as 3D one by using level set approach to handle the topology changes such as splitting and merging the contours. For the proposed ROI segmentation algorithm, one of the geometric deformable models, active contours without edges, is used. This technique is capable of segmenting the regions with either weak edges, or even, no edges at all. The results of the proposed ROI segmentation algorithm are compared with those of the two experts' manual marking. The results are also compared with the common regions manually marked by both experts and with the total regions marked by either expert. The proposed ROI segmentation algorithm is also evaluated by using region-based and pixel-based strategies. The evaluation results indicate that the proposed algorithm produces similar results to those of the experts' manual markings, but with the added advantages of being fast and reliable. This novel algorithm also detects some regions that have been missed by one expert but confirmed by the other. In conclusion, the two newly devised algorithms can assist experts in segmenting the prostate image and detecting the suspicious abnormal regions that should be considered for biopsy. This leads to the reduction the number of biopsies, early detection of the diseased regions, proper management, and possible reduction of death related to prostate cancer.

Im Rahmen einer prospektiven Studie wurden zwischen Juli 2007 und Oktober 2008 Patienten mit Prostatakarzinom in der Klinik für Urologie des Universitätsklinikums Leipzig befragt. Die Lebensqualität und Lebenszufriedenheit wurden mit Hilfe von Selbstbeurteilungsfragebögen (EORTC QLQ-C30, SF-8 und FLZM) erfasst. Insgesamt lagen Daten von 276 Patienten vor. Ziel der Arbeit war es, den Verlauf von Lebensqualität und Lebenszufriedenheit über drei Monate nach dem Klinikaufenthalt zu beobachten, sowie diese Daten mit denen der Allgemeinbevölkerung zu vergleichen. Weiterhin wurden verschiedene Einflussfaktoren auf Lebensqualität und Lebenszufriedenheit bewertet. Prostatakarzinompatienten gaben die schlechteste Lebensqualität zum Zeitpunkt vierzehn Tage nach Entlassung aus der Klinik an. Innerhalb von drei Monaten erreichten sie wieder das Ausgangsniveau an Lebensqualität. Insgesamt zeigten sich kaum klinisch bedeutsame Unterschiede in der Lebensqualität zwischen Patienten und Allgemeinbevölkerung. Die Lebenszufriedenheit nahm im Verlauf zwar ab, jedoch gaben die Prostatakrebspatienten zu fast allen Zeitpunkten eine ähnliche oder sogar höhere Lebenszufriedenheit als die Vergleichsgruppe an. Die einzige, aber wichtige Ausnahme hiervon bildete der Bereich Sexualität, hier waren die Patienten nach 3 Monaten deutlich unzufriedener als die Männer der Allgemeinbevölkerung. Alter, Bildungsgrad und die seit Diagnosestellung vergangene Zeit stellten sich im Gegensatz zum Tumorstadium als Einflussfaktoren für die Beurteilung der Lebensqualität dar. Alle diese Faktoren zeigten jedoch keinen signifikanten Einfluss auf deren Lebenszufriedenheit. Die Korrelationen zwischen den drei eingesetzten Fragebögen waren fast ausnahmslos positiv, entsprechend einer gleichsinnigen Variabilität. Eine generelle psychoonkologische Betreuung aller Patienten scheint nicht notwendig, jedoch sollten Ärzte für die Probleme des Einzelnen sensibilisiert sein um rechtzeitig Hilfe anbieten zu können.

<p>Abstract</p><p>The trefoil factor family of secreted proteins contains three members; trefoil factor 1 or TFF1, trefoil factor 2 or TFF2, and trefoil factor 3 or TFF3. These three proteins share a conserved 42-43 amino acid domain containing 6 cysteine residues resulting in three disulfide bonds that holds the protein in a characteristic three-loop or "trefoil structure" known as the P domain. TFF1 is primarily localized to the stomach and secreted by the gastric mucosa while TFF2 and TFF3 are primarily localized to the colon and duodenum and secreted by the goblet cells. All three of these proteins play a protective role in the gastrointestinal tract where they are normally localized and have been identified as possible tumor suppressors, however, these proteins are also upregulated in cancer within tissues where they are not normally expressed including the breast, pancreas, prostate, and liver. The mechanisms by which two of these factors, TFF1 and TFF3, promote tumorigenesis remain largely undefined. In this dissertation we will attempt to elucidate these mechanisms as well as the regulation of these two proteins in both pancreatic and prostate cancer. Many of the underlying genetic and molecular mechanisms involved in the development of both pancreatic and prostate cancer remain largely unknown and as a result, therapeutic and diagnostic tools for treating these diseases are not as effective as they could be. By deciphering the role of TFF1 and TFF3 in these cancers, they could potentially serve as new therapeutic targets or biomarkers for treating both diseases.</p><p>Chapter 2 of this dissertation will examine the functional role of TFF1 promoting tumorigenesis in pancreatic and prostate cancer. We will show that TFF1 expression is critical for the viability of both pancreatic and prostate cancer cells and that reduction of TFF1 expression in these cells results in decreased tumorigenicity when implanted in immunocompromised mice. It will also be demonstrated that TFF1's function in promoting tumorigenicity is its ability to assist tumor cells overcome the tumor suppressive barrier of senescence. Thirdly, we show that the form of senescence that TFF1 assists in allowing the cells overcome is oncogene-induced senescence (OIS). Lastly, a cell cycle array identifies the potential downstream target p21CIP, a cyclin-dependent kinase inhibitor and OIS marker, whose expression is induced by loss of TFF1 expression.</p><p>In Chapter 3 of this work, we examine the role of another trefoil factor family member, TFF3, and its role in promoting prostate tumorigenesis. Just as with TFF1, it appears that TFF3 3 expression is critical for prostate cancer cell viability and tumorigenicity using the same experimental techniques used in Chapter 2. Using a genetically defined model of prostate cancer, a PI3-kinase-dependent regulatory mechanism of TFF3 emerges in this prostate cancer context. Using this system we begin to see a divergence in both regulation and function of TFF1 and TFF3 in prostate cancer. Finally, a mouse model expressing TFF3 was developed to monitor the histopthological changes associated with expression of this protein. Initial characterization of this model suggests a hyperplastic phenotype coinciding with TFF3 expression in the prostate.</p><p>The two studies in this dissertation establish a role of TFF1 and TFF3 in both prostate and pancreatic tumorigenesis and demonstrate that ablation of expression of both proteins is a potent inhibitor of tumorigenesis. With this knowledge, it is possible that TFF1 and TFF3 may become a potential therapeutic target or diagnostic marker for better treatment of prostate and pancreatic cancer.</p> / Dissertation

<p>Glucose metabolism remains an intensely explored topic of cancer biology since the initial discoveries of Otto Warburg nearly 80 years ago. Many solid tumors metabolize glucose primarily to lactate despite the availability of oxygen, revealing a dependence on glycolysis that may serve as a basis for targeted therapy. In particular, a diet devoid of carbohydrate may minimize the growth capabilities of glucose-dependent cancers. As our interests lie in prostate cancer, we examined whether a ketogenic diet devoid of carbohydrate (NCKD) would reduce the growth rate of tumors derived from human prostate cancer cell lines in a murine xenograft model.</p> <p>Our initial experiments utilized the LAPC-4 cell line, a human androgen-sensitive prostate cancer cell line, in a SCID-mouse xenograft model to determine the effects of an NCKD on tumor growth and animal survival relative to two other diets: (1.) a Western-type diet (WD) reflecting consumptions patterns of men diagnosed with prostate-cancer in the Western world and (2.) a low-fat diet (LFD) representing the present standard of care. Following this study, we conducted a second study utilizing a different human prostate cancer cell line (LNCaP) in order to assess whether our initial observations were robust across multiple prostate cancer tumor models and to also further explore the molecular underpinnings of our observations. Both studies revealed the NCKD leads to a reduction in tumor growth rate and greater overall mouse survival relative to the WD. In addition, the NCKD was equivalent in these parameters to the LFD. We also observed key associations between survival and extent of urinary ketosis as well as favorable changes in insulin and insulin-like growth factor-1 (IGF-1) and gene expression that would be predictive of prolonged survival in mice consuming the NCKD.</p> <p>We believe these data provide compelling evidence to consider a potential therapeutic role for dietary carbohydrate restriction in prostate cancer. We hope these results ultimately serve as a basis to conduct future clinical trials assessing whether dietary carbohydrate restriction, either alone or in combination with more conventional therapies, provides clinicians with an additional weapon against prostate cancer.</p> / Dissertation