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Psychopathic personality traits are positively associated with externalising behaviours, and negatively associated with internalising behaviours. However, the contribution of different facets of psychopathy (boldness, meanness, disinhibition) in explaining externalising and internalising behaviours across genders are inconsistent. In this study, we explored gender differences in the assessment of, and relationships between, psychopathic personality traits, trait anxiety, social anxiety, depression, mental health, and aggressive behaviour in 822 students from a German University (586 women, 236 men; Mage = 22.27). Using a structural equation model, we found a positive relationship between aggressive behaviour and all three facets of psychopathy, a positive relationship between internalising behaviours and disinhibition, and a significant negative relationship between internalising behaviours and boldness. Despite gender differences in absolute levels of these variables, the overall pattern of the relationships between variables was consistent across genders. This indicates that symptom level differences across gender cannot be accounted for by variation in early developing personality traits like psychopathy.

253 Background: Perineural Invasion (PNI) is defined as malignant epithelial cell invasion of the perineural space and nerves. Despite widespread acknowledgement of the clinical significance of PNI as a PCa pathological finding associated with recurrence, increased risk of bone metastasis and poor survival, the molecular mechanism underlying this pathology is relatively unknown. The malignant epithelial cells within the PNI potentially provides a spatially defined “snapshot” of disease progression, as the cells switch to a more migrational phenotype associated with metastatic progression. Here we present the initial spatial PNI phenotypic characterisation in PCa. Methods: Archival FFPE blocks, with associated full clinical history, from patients who underwent a radical prostatectomy for prostate cancer were retrieved under research ethics REC#07/H1003/161+5 10_NOCL_02. Biomarkers EphA2, pEphA2s897, pMLC2, E-Cadherin, Vimentin, TOMM20, MTC01, NDUFB8, PTEN were assessed on 4µm serial sections stained using a multiplex TSA protocol, with S100, pan-cytokeratin and DAPI acting as landmarks, on a Ventana Discovery platform prior to scanning on a Versa 3 platform with Halo image analysis. Prostate zones were defined at 500µm intervals either side of the prostate capsule. Univariate and multivariate (hierarchical clustering, UMap clustering) expression analysis and correlation with clinic-pathological features was conducted within R. Results: The PNI epithelial cells within each spatial zone of the prostate are significantly different to each other (Kruskal-Wallis test p < 2.2x10−16 except for MTC01 p = 5.3x10−10). In comparison with the local tumour lesion, PNI epithelial cells localised within 1000µm of the prostate edge and outside the tumour lesion, have undergone a migrational switch, gaining features associated with an activated metastatic phenotype, with increased expression of amoeboid signalling (EphA2, pEphA2s897, pMLC2) and mitochondrial defects (loss of Complex I and IV, gain of mitochondrial mass (TOMM20)). Patients clustering by multivariate expression trends across the prostate regions showed 4 distinct patient groups, with PNI epithelial cells in patient group 1 & 2 displaying a more epithelial to mesenchymal (EMT) phenotype, especially in the first 1000µm inside the prostate organ. Conclusions: Cells within PNI close to the edge of the prostate have features consistent with a switch to migrational/metastatic activation in contrast to the more indolent cell type found deeper within the tumour. Further characterisation of this localised migrational upregulation will help in understanding the transition from a localised to a metastatic phenotype.

Background Docetaxel chemotherapy is a standard of care for metastatic castrate-resistant prostate cancer (mCRPC): 40–50% of patients achieve a biochemical response. However, there is a lack of response predictive biomarkers. Objective To assess lactate dehydrogenase (LDH) as a docetaxel response biomarker in mCRPC and to examine the association of LDH with genomic alterations in primary diagnostic biopsies. Design, setting, and participants Clinical and associated primary tumour-targeted next-generation sequencing data from matched training (n = 150) and test (n = 120) cohorts of progressive mCRPC patients receiving docetaxel therapy were analysed. Data were correlated with large-scale prostate cancer genomic datasets. Outcome measurements and statistical analysis Prostate-specific antigen (PSA) response, radiographic response, biochemical progression-free survival (PFS), overall survival (OS), genomic analysis of primary biopsies, and genomic datasets (Memorial Sloan Kettering Cancer Center [MSKCC] and SU2C/PCF). Results and limitations Serum LDH ≥450 U/l is a reliable prognostic biomarker (area under the curve: 0.757 [standard deviation 0.054, 95% confidence interval [CI] 0.650–0.864, p < 0.001]) in progressive mCRPC, predicting PFS at 3 mo. Patients with LDH ≥450 U/l were poorer PSA responders, with shorter PFS (213 vs 372 d, hazard ratio [HR] 1.876, 95% CI 1.289–2.7300) and OS (362 vs 563 d, HR 1.630, 95% CI 1.127–2.357). High LDH is an independent surrogate marker for survival following docetaxel and predicts a poor radiological response (p = 0.043). Of the 14 patients with LDH ≥450 U/l available for next-generation sequencing, nine (64.3%) were more likely to have DNA repair gene mutation(s) (BRCA1/2, ATM, CHEK2, Fanconi anaemia gene) in their primary biopsy. Cross correlation with MSKCC and SU2C/PCF databases revealed a positive correlation between LDHA, PARP1 (r = 0.667, p < 0.01), and other DNA repair genes. Conclusions Genomic abnormalities of LDHA and DNA repair in primary biopsies link to high pretreatment LDH and poor response to docetaxel in mCRPC. Patient summary The presence of mutations of the lactate dehydrogenase and DNA repair pathways are associated with aggressive prostate cancer and poor response to chemotherapy later in the disease.

The Notch ligand Jagged1 is subject to regulated intramembrane proteolysis (RIP) which yields a soluble ectodomain (sJag) and a soluble Jagged1 intracellular domain (JICD). The full-length Jagged1 protein enhances prostate cancer (PCa) cell proliferation and is highly expressed in metastatic cells. However, little is known regarding the mechanisms by which Jagged1 or its RIP-generated fragments might promote PCa bone metastasis. In the current study we show that bone marrow stroma (BMS) induces Jagged1 expression in bone metastatic prostate cancer PC3 cells and that this enhanced expression is mechanistically linked to the promotion of cell migration. We also show that RIP-generated Jagged1 fragments exert disparate effects on PC3 cell behaviour and Notch signaling. In conclusion, the expression of both the full-length ligand and its RIP-generated fragments must be considered in tandem when attempting to regulate Jagged1 as a possible PCa therapy.

A disintegrin and metalloproteinases (ADAMs) and matrix metalloproteinases (MMPs) are zinc metalloproteinases (ZMPs) that catalyze the “ectodomain shedding” of a range of cell surface proteins including signaling and adhesion molecules. These “sheddases” are associated with the invasion and metastasis of a range of cancers. Increased serum and tumor tissue levels of copper are also observed in several cancers, although little is known about how the metal might promote disease progression at the molecular level. In the current study, we investigated whether copper might regulate the ectodomain shedding of two key cell surface proteins implicated in the invasion and metastasis of prostate cancer, the Notch ligand Jagged1 and the adhesion molecule E-cadherin, and whether the metal was able to influence the invasion of the prostate cancer epithelial cell line PC3. Physiological copper concentrations stimulated the ZMP-mediated proteolysis of Jagged1 and E-cadherin in cell culture models, whereas other divalent metals had no effect. Copper-mediated Jagged1 proteolysis was also observed following the pretreatment of cells with cycloheximide and in a cell-free membrane system, indicating a posttranslational mechanism of sheddase activation. Finally, the concentrations of copper that stimulated ZMP-mediated protein shedding also enhanced PC3 invasion; an effect that could be negated using a sheddase inhibitor or copper chelators. Collectively, these data implicate copper as an important factor in promoting prostate cancer cell invasion and indicate that the selective posttranslational activation of ZMP-mediated protein shedding might play a role in this process. Mol Cancer Res; 10(10); 1282–93. ©2012 AACR.

Background: Although statins do not affect the incidence of prostate cancer (CaP), usage reduces the risk of clinical progression and mortality. Although statins are known to downregulate the mevalonate pathway, the mechanism by which statins reduce CaP progression is unknown. Methods: Bone marrow stroma (BMS) was isolated with ethical approval from consenting patients undergoing surgery for non-malignant disease. PC-3 binding, invasion and colony formation within BMS was assessed by standardised in vitro co-culture assays in the presence of different statins. Results: Statins act directly on PC-3 cells with atorvastatin, mevastatin, simvastatin (1 μℳ) and rosuvastatin (5 μℳ), but not pravastatin, significantly reducing invasion towards BMS by an average of 66.68% (range 53.93–77.04% P

Which people are most swayed by self-image motives and hence most likely to make consumer choices in line with those motives? This article contends that the answer is narcissists —individuals who see themselves, and who want others to see them, as special, superior, and entitled and who are prone to exhibitionism and vanity. This work hypothesizes that narcissists will, to validate their excessively positive self-views, strive to purchase the high-prestige products (i.e., expensive, exclusive, new, and flashy). In so doing, they will regulate their own esteem by increasing their apparent status and consequently earning others’ admiration and envy. This article also hypothesizes that narcissists will show greater interest in the symbolic than utilitarian value of products and will exhibit, even controlling for self-esteem, more pronounced self-enhancement phenomena such as endowment and self-signaling effects.

Objectives: We introduce biochemistry as a second dimension to Gleason grading, using Fourier transform infrared (FTIR) microspectroscopy. For the first time, we correlate FTIR spectra derived from prostate cancer (pCA) tissue with Gleason score and the clinical stage of the tumour at time of biopsy. Methods: Serial sections from paraffin-embedded pCA tissue were collected. One was stained with hematoxylin and eosin and Gleason scored; FTIR spectra were collected from malignant locations using a second unstained section. FTIR spectra, representing different Gleason grades, were used to construct a diagnostic classifier for pCA using linear discriminant analysis (LDA). This model was blind tested using 383 IR spectra from 36 biopsies. Results: Using a three-band Gleason criteria, we obtained sensitivity of ≥70% for the FTIR-LDA model to predict Gleason 7, with specificities of ≥81%. Using a threshold of Gleason/FTIR-LDA score of ≥8, we obtained a sensitivity and specificity of 71% and 67%, respectively, for the correlation with metastatic tumours using the FTIR-LDA system and 85% and 63%, respectively, for the correlation of metastatic tumours using the Gleason system. Conclusions: There is a correlation between tissue architecture using Gleason score with tissue biochemistry using FTIR-LDA. Both systems are similar in their performance in predicting metastatic behaviour in tumours from individual patients. © 2006 European Association of Urology.

Epidemiological studies have shown not only a relationship between the intake of dietary lipids and an increased risk of developing metastatic prostate cancer, but also the type of lipid intake that influences the risk of metastatic prostate cancer. The Omega-6 polyunsaturated fatty acid, Arachidonic acid, has been shown to enhance the proliferation of malignant prostate epithelial cells and increase the risk of advanced prostate cancer. However, its role in potentiating the migration of cancer cells is unknown. Here we show that arachidonic acid at concentrations ≤5 μM is a potent stimulator of malignant epithelial cellular invasion, which is able to restore invasion toward hydrocortisone-deprived adipocyte-free human bone marrow stroma completely. This observed invasion is mediated by the arachidonic acid metabolite prostaglandin E2 and is inhibited by the Omega-3 poly-unsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid at a ratio of 1:2 Omega-3:Omega-6, and by the COX-2 inhibitor NS-398. These results identify a mechanism by which arachidonic acid may potentiate the risk of metastatic migration and secondary implantation in vivo, a risk which can be reduced with the uptake of Omega-3 poly-unsaturated fatty acids. © 2006 Cancer Research UK.

Background Prostate stem cells, responsible for the development, maturation, and function of the prostate, have been implicated in the aetiology of both benign prostate hyperplasia (BPH) and prostate cancer (CaP). However, research has been hampered by the lack of a definitive stem cell marker. We have adapted the protocol for differential Hoechst 33342 uptake by hemopoietic stem cells to enable isolation of putative stem cells from the prostate. Methods Prostate epithelial cells isolated from prostate tissue obtained from patients with BPH after transurethral resection of the prostate were stained with Hoechst 33342. The Hoechst 33342 Red/Blue flow cytometry profile was then determined. Hoechst 33342 and Pyronin Y staining was used to determined the cell cycle status. Results A verapamil-sensitive side population (SP) can be isolated from primary prostate tissue accounting for 1.38% ± 0.07% of prostate epithelial cells. Cell cycle analysis of this SP population revealed that the majority of SP cells are in either G0 (12.38 ± 0.31%) or G1 (63.19 ± 2.13%). Conclusions The Hoechst 33342 dye efflux protocol can be adapted for the isolation of a SP from primary prostate tissue. Cytometry Part A 54A:89–99, 2003. © 2003 Wiley-Liss, Inc.

BACKGROUND. The metastatic potential of a series of prostate cell lines was analysed bymeasuring motility and invasiveness, and further correlated to the expression of epithelialdifferentiation markers.METHODS. Invasion and motility were measured using in vitro assays. Immunohistochem-istry of cell lines and tissues was used to identify expression of cytokeratins 8 and 1, 5, 10, 14,vimentin, prostate specific antigen, prostate specific membrane antigen, androgen receptor,desmoglein, E-cadherin, b1 integrin, CD44, hmet, vinculin and actin.RESULTS. Expressionofvimentinwastheonlymarkertocorrelatewithmotility, nomarkerscorrelated to invasion. Lower vimentin expression was observed in cells with low motility(PNT2-C2) and high expression in cells with high motility (P4E6, PNT1a, PC-3). Vimentinexpression was not detected in well differentiated tumours, moderately differentiated tumorscontained vimentin positive cells (1/9 bone scan negative, 2/5 bone scan positive), but themajority of poorly differentiated cancers (4/11 bone scan negative, 9/14 bone scan positive)and bone metastases (7/8) had high vimentin expression in tumor cells.CONCLUSIONS. Motile prostate cancer cell lines express vimentin. In tissue sections, thepresence of vimentin positive tumour cells correlated positively to poorly differentiatedcancersandthepresenceofbonemetastases. Prostate52:253–263,2002.

Empathy plays a critical role in fostering and maintaining social relations. Narcissists lack empathy, and this may account for their interpersonal failures. But why do narcissists lack empathy? Are they incapable, or is change possible? Three studies addressed this question. Study 1 showed that the link between narcissism and low empathy generalizes to a specific target person presented in a vignette. The effect was driven by maladaptive narcissistic components (i.e., entitlement, exploitativeness, exhibitionism). Study 2 examined the effect of perspective-taking (vs. control) instructions on self-reported responses to a video. Study 3 examined the effect of the same manipulation on autonomic arousal (heart rate [HR]) during an audio-recording. Perspective-taking ameliorated negative links between maladaptive narcissism and both self-reported empathy and HR. That is, narcissists can be moved by another’s suffering, if they take that person’s perspective. The findings demonstrate that narcissists’ low empathy does not reflect inability, implying potential for intervention.

We review the literature on the relation between narcissism and consumer behavior. Consumer behavior is sometimes guided by self-related motives (e.g., self-enhancement) rather than by rational economic considerations. Narcissism is a case in point. This personality trait reflects a self-centered, self-aggrandizing, dominant, and manipulative orientation. Narcissists are characterized by exhibitionism and vanity, and they see themselves as superior and entitled. To validate their grandiose self-image, narcissists purchase high-prestige products (i.e., luxurious, exclusive, flashy), show greater interest in the symbolic than utilitarian value of products, and distinguish themselves positively from others via their materialistic possessions. Our review lays the foundation for a novel methodological approach in which we explore how narcissism influences eye movement behavior during consumer decision-making. We conclude with a description of our experimental paradigm and report preliminary results. Our findings will provide insight into the mechanisms underlying narcissists’ conspicuous purchases. They will also likely have implications for theories of personality, consumer behavior, marketing, advertising, and visual cognition.\ud \ud “Some people think luxury is the opposite of poverty. It is not. It is the opposite of vulgarity.”\ud \ud – Coco Chanel

Background:High intake of omega-6 polyunsaturated fatty acids (PUFA) has been associated with clinical progression in prostate cancer (CaP). This study investigates the signalling mechanism by which the omega-6 PUFA arachidonic acid (AA) induces prostatic cellular migration to bone marrow stroma.Methods:Western blot analysis of the PC-3, PC3-GFP, DU 145 and LNCaP cells or their lipid raft (LR) components post AA stimulation was conducted in association with assays for adhesion and invasion through the bone marrow endothelial monolayers.Results: Arachidonic acid increased transendothelial migration of PC3-GFP cells (adhesion 37%±0.08, P0.0124; transmigration 270%±0.145, P0.0008). Akt, Src and focal adhesion kinase (FAK) pathways were induced by AA and integrally involved in transendothelial migration. LR were critical in AA uptake and induced Akt activity. Ephrin receptor A2 (EphA2), localised in LR, is expressed in DU 145 and PC-3 cells. Arachidonic acid induced a rapid increase of EphA2 Akt-dependent/ligand-independent activation, while knockdown of the EphrinA1 ligand decreased AA induced transendothelial migration, with an associated decrease in Src and FAK activity. Arachidonic acid activated Akt in EphA2 LNCaP cells but failed to induce BMEC transendothelial invasion.Conclusion:Arachidonic acid induced stimulation of EphA2 in vitro is associated fundamentally with CaP epithelial migration across the endothelial barrier. © 2012 Cancer Research UK. All rights reserved.

INTRODUCTION AND OBJECTIVES: Exposure of cancer cells to statins result in the down-regulation of the mevalonate pathway by HMG-CoA reductase inhibition, inducing inhibition of a range of cellular functions such as membrane integrity, cell signalling, protein synthesis and cell cycle progression. Disruption of these processes may result in changes in the regulation of tumour initiation, growth and metastasis. Clinical studies have reported that statins do not effect the incidence of clinically significant prostate cancer but usage reduces the risk of progression and mortality (0.26; 95% CI 0.08–0.83 0.64; 95% CI 0.44–0.93). It is currently unclear how statins reduce the clinical progression of prostate cancer. Here we present in vitro data showing that statins act directly on the prostate epithelial cells, reducing their ability to invade through and form metastatic colonies in the bone marrow stroma (BMS). METHODS: BMS was isolated with ethical approval from consenting patients undergoing surgery for non-malignant disease. All statins were used at non-toxic doses as determined by colony forming assays (atorvastatin, mevastatin, simvastatin (1 M), rosuvastatin (5 M) & pravastatin (100 M)). Binding of statin treated PC-3GFP cells to BMS was assessed using a FLUOstar OPTIMA spectrometer. Invasion of statin treated PC-3 cells towards untreated BMS or 7 day statin pre-treated BMS was assessed using the FluoroBlok tumour co-culture cell invasion system. Colony formation was assessed by seeding statin treated PC-3 cells on to confluent BMS. Statins were refreshed daily for 8 days before fixing and staining with anti pancytokeratin, counting and measuring epithelial colonies. RESULTS: Statins do not affect the ability of PC-3 cells to bind to BMS. Pravastatin had no effect on PC-3 invasion or colony formation in BMS. Treating PC-3 cells with atorvastatin, mevastatin, simvastatin and rosuvastatin significantly reduced their ability to invade towards BMS as compared with untreated controls on average 66.68% (range 53.93%–77.04% p 0.05). These statins also significantly reduced the number (76.2 vs 118; p 0.05) and size (0.2mm vs 0.29mm; p 0.05) of colonies formed in BMS co-culture, with colonies displaying a more compact morphology indicative of a reduction in the ability to migrate through the BMS. CONCLUSIONS: Statins reduce the risk of prostate cancer progression by inhibiting the ability of malignant prostate epithelial cells to invade towards and through human BMS, reducing the formation and the spread of metastatic prostate colonies.

It is hypothesized that cells with stem cell-like properties may be influential in carcinogenesis, possessing the ability to self-renew, produce differentiated daughter cells and resist environmental or therapeutic injury. This has led to a surge in interest in identifying and characterizing the tumour initiating or cancer stem cell (CSC) with the aim of discovering novel diagnostic and prognostic markers and of understanding the basic biology with the ultimate aim of generating new therapeutic approaches and biomarkers. However, a major hurdle to this process has been the lack of a truly specific cancer stem cell biomarker allied to the rarity of these cells. This has led to problems in characterising these CSCs by traditional '-omic' techniques. Using a renal carcinoma cell line model, we show that synchrotron radiation-Fourier transform infrared (SR-FTIR) spectroscopy is a suitable tool to measure discrete differences in the biochemistry of small numbers of single-cells. Using the chemometric techniques of Principal Component and Linear Discriminant Analysis (PCA and LDA) for multivariate reduction, biochemical differences between the cells from different sub-populations were evaluated. Results found lipid and phosphodiester vibrations to be particularly good discriminating markers in the spectra of these stem-like cells, relative to the more differentiated, proliferating cells that make up the majority of the cell population. © 2010 The Royal Society of Chemistry.

Interest in developing robust, quicker and easier diagnostic tests for cancer has lead to an increased use of Fourier transform infrared (FTIR) spectroscopy to meet that need. In this study we present the use of different experimental modes of infrared spectroscopy to investigate the RWPE human prostate epithelial cell line family which are derived from the same source but differ in their mode of transformation and their mode of invasive phenotype. Importantly, analysis of the infrared spectra obtained using different experimental modes of infrared spectroscopy produces similar results. The RWPE family of cell lines can be separated into groups based upon the method of cell transformation rather than the resulting invasiveness/aggressiveness of the cell line. The study also demonstrates the possibility of using a genetic algorithm as a possible standardised pre-processing step and raises the important question of the usefulness of cell lines to create a biochemical model of prostate cancer progression. © 2010 The Royal Society of Chemistry.

Background: Prostate cancer (CaP) preferentially metastasises to the bone, and we have previously shown that the poly-unsaturated fatty acid (PUFA) arachidonic acid (AA) is a potent stimulator of CaP invasion. Here we present that AA promotes CaP invasion by inducing bone marrow adipocyte formation. Methods: Boyden invasion-chamber assays assessed the ability of dietary oils, their PUFA components, and specific PUFA-loaded adipocytes to induce PC-3 invasion. Lipid transfer and metabolism was followed using deuterated AA and Fourier Transform Infrared spectroscopy (FTIR). Results: Poly-unsaturated fatty acid constituents, but not their corresponding dietary oils, induced PC-3 invasion. PUFAs induce bone marrow adipocyte (BM-Ad) differentiation with AA inducing higher levels of BM-Ad differentiation, as compared with other PUFAs (3998514.4 vs 932265.8; P0.00002), which stimulated greater PC-3 invasion than free AA (22 408.5607.4 vs 16 236313.9; P0.01111) or adipocytes generated in the presence of other PUFAs. In bone marrow co-culture PC-3 and BM-Ad interactions result in direct uptake and metabolism of AA by PC-3 cells, destruction of the adipocyte and subsequent formation of a bone metastasis. Conclusion: The data supports the hypothesis that AA not only promotes CaP invasion, it also prepares the soil, making it more supportive for implantation and propagation of the migrating metastatic cell. © 2010 Cancer Research UK. All rights reserved.

Prostate cancer (PCa) preferentially metastasizes to the bone marrow stroma of the axial skeleton. This activity is the principal cause of PCa morbidity and mortality. The exact mechanism of PCa metastasis is currently unknown, although considerable progress has been made in determining the key players in this process. In this review, we present the current understanding of the molecular processes driving PCa metastasis to the bone.

BACKGROUND: The prostate epithelial stem cell has been proposed as the primary origin of neoplastic change in prostate cancer. However, the isolation and characterization of unexpanded prostate epithelial stem cells have proven problematic. METHODS: A prostate epithelial side population (SP) has been isolated utilizing a modified Hoechst 33342 dye efflux assay from both benign and malignant prostate tissue. CD45(-ve), integrin alpha2(+ve) Hoechst 33342 SP and NSP cells were isolated by FACS, immunophenotyped and functionally characterized in 3D culture. RESULTS: FACS analysis revealed a verapamil sensitive SP accounting for 0.93 +/- 0.12% and 0.57 +/- 0.11% of the total epithelial population from both benign and malignant prostates. The benign SP phenotype revealed a heterogeneous cell population consisting predominantly of small basal cells containing minimal cytoplasm. Conversely, the malignant SP was of undetermined acinar origin and with a complete loss of expression of the CDK2 inhibitor p21(WAF1/Cip1). In vitro androgen-enhanced 3D culture of the benign and malignant SP cells led to the production of spheroids which had acinus like morphology and expressed primitive and basal cell markers. Incorporation of the CD133 marker isolated a further SP sub-fraction accounting for 0.037 +/- 0.01% of epithelial cells. CONCLUSIONS: Our observations are consistent with the Hoechst 33342 dye efflux assay isolating a stem cell enriched population which can be further sub-fractionated by CD133 selection. Moreover, the loss of the CDK inhibitor in malignancy is consistent with the hypothesis that neoplastic change originates in the stem cell compartment. Prostate 67: 1384-1396, 2007. (c) 2007 Wiley-Liss, Inc.

Prostate cancer (CaP) cells preferentially metastasise to the bone marrow, a microenvironment that plays a substantial role in the sustenance and progression of the CaP tumour. Here we use a combination of FTIR microspectroscopy and histological stains to increase molecular specificity and probe the biochemistry of metastatic CaP cells in bone marrow tissue derived from a limited source of paraffin-embedded biopsies of different patients. This provides distinction between the following dominant metabolic processes driving the proliferation of the metastatic cells in each of these biopsies: glycerophospholipid synthesis from triacylglyceride, available from surrounding adipocytes, in specimen 1, through significantly high (p

Introduction and Objective: Metastatic progression of prostate cancer requires a change of state, epithelial to mesenchymal transition (EMT), reducing cell to cell adherence and contact, with associated cell motility and invasion. EMT is one of the hallmarks of cancer and characterisation of the EMT process has defined a panel of EMT biomarkers. Here we investigate the potential of an EMT panel of biomarkers in an unscreened population at first presentation with prostate cancer. Methods: A prostate Tissue Micro Array (TMA) comprising 2,450 cores containing normal adjacent, prostatic intraepithelial neoplasia and malignant prostate tissue from 524 diagnostic biopsies, linked to an extensive clinical database with long-term outcome, was studied. Expression of a panel of EMT biomarkers, including E-cadherin and vimentin, was assessed using an automated, high throughput fluorescent staining (Leica BOND-MAX), image capture (Carl Zeiss Microimaging Mirax scanner) and quantitative image analysis (Definiens Tissue Studio) protocol. Definiens H-Score was converted to pathological scores prior to univariate and multivariate Cox proportional hazards regression analysis in SPSS. Results: Expression of an EMT profile predicted a significant reduction in both overall and cancer specific survival. In univariate analysis, low or negative E-cadherin staining was associated with a reduction in median overall survival by 45 months (95% CI: 10.4-79.6; P Conclusions: Loss of E-cadherin and concomitant expression of the EMT marker vimentin are powerful predictors of prostate cancer outcome at first presentation. Citation Format: Ahmad Al-Sukaini, Claire Hart, Richard Robinson, Mick Brown, Noel Clarke. Prognostic potential of epithelial to mesenchymal transition in prostate cancer at first presentation. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4350. doi:10.1158/1538-7445.AM2015-4350

The natural history of prostate cancer is notoriously variable and whilst some tumours remain indolent for many years, others result in rapid progression to metastasis and death. A substantial minority of patients treated with radical prostatectomy or radiotherapy with curative intent have rising levels of PSA in their serum after treatment, and this is often associated with the eventual development of metastases. It is possible that some of these patients already have ‘micrometastases’ at the time of treatment. No staging investigation is able to predict reliably which patients will be cured by radical treatment. Radionuclide bone scans are positive in only 5% of patients with a serum PSA of

Objectives: To determine the mode of action of Zoledronic acid in the inhibition of metastasis in prostate cancer and the reduction of prostate cancer bone metastases. Methods: Benign and malignant primary prostatic epithelial cells (PEC) and the PC-3 prostate cancer cell line were studied in co-culture using human bone marrow stroma in the presence of escalating doses of EDTA, Clodronate, Pamidronate and Zoledronic acid. PEC binding and colony growth in bone marrow stroma was measured using standardised quantitative techniques. PEC cellular invasion through Matrigel and an endothelial monolayer was measured either in invasion chambers or by the measurement of endothelial monolayer permeability to fluorescent dextran. Co-culture supernatants were assayed for specific cytokine levels. Bone marrow cellular toxicity was assessed using a standard Mix assay. Results: Treatment of PEC with up to 100 μM bisphosphonate did not affect their ability to bind to bone marrow endothelium or stroma. Bone marrow endothelial permeability was reduced by 100 μM Zoledronic acid by 3.8% (p=0.03856). Both Pamidronate (40% at 100 μM, p≤0.05) and Zoledronic acid inhibited PEC invasion, with Zoledronic acid being the most potent (40% at 10 μM, p≤0.05 rising to 91% at 100 μM, p≤0.001). Zoledronic acid inhibits malignant PEC proliferation in bone marrow stroma co-culture (26.5% at 10 μM rising to 66.5% at 40 μM). This was accompanied by changes within the cytokine milieu with a >800% rise in TIMP-2. Conclusion: Zoledronic acid is a potent inhibitor of PEC invasion across bone marrow endothelium and colony formation with the bone marrow stroma, affecting the MMP: TIMP-2 balance to favour MMP inhibition. © 2004 Elsevier B.V. All rights reserved.

Imaging ToF-SIMS and synchrotron-based Fourier transform infrared (SR-FT-IR) microspectroscopy have been used to obtain chemical information from individual cells derived from human prostate cancer (CaP). ToF-SIMS imaging of molecular signals characteristic of membrane bound phospholipids are used to elucidate different fracture planes within individual freeze-fractured CaP cells. The localisation of Cu within the cytoplasm of cancer cells is consistent with increased metastatic potential. Line scans across CaP cells using SR-FT-IR micro spectroscopy provide complimentary information on the localisation (+/-1 mum) of lipid and protein domains. This combined analytical approach offers a novel means of characterising individual CaP cells and investigating the biochemical basis of disease progression and metastases. (C) 2004 Elsevier B.V. All rights reserved.

181 Background: Tissue microarrays (TMAs), linked to clinical databases containing clinical outcome data, offer a great potential for defining and validating new biomarkers in urological oncology. The long term follow up required for true clinical outcome prostate cancer results in the use of archival material for TMA construction. The use of such material has inherent problems which must be overcome to unlock the full potential of prostate TMAs. Methods: A prostate 2,450 core TMA containing normal, prostatic intraepithelial neoplasia and malignant prostate tissue from 524 archival patient specimens was constructed and analysed. Each core was independently verified histologically. An automated, high throughput fluorescent staining (Leica BOND-MAX), image capture (Carl Zeiss Microimaging Mirax scanner) and scoring (Definiens Tissue Studio) protocol was compared with standardised conventional staining and manual scoring techniques. Results: Image analysis showed excellent correlation (r=0.785) and absolute agreement between the automated and manual scoring protocols (ICC=0.769, 95% CI [0.629-0.877]). Replicate analysis demonstrated that the automated, high throughput protocol was more reliable and consistent than the standardised manual staining and scoring protocols. The increased sensitivity of fluorescent staining in the automated protocol highlighted significant (p>0.0001) antigen degradation (pan-cytokeratin), hence poorly persevered regions, in 50.7% of the archival TMA cores. Using well preserved cores the potential of combining Erg expression with EMT markers as a prognostic marker will be presented. Conclusions: This study shows that archival TMAs can be utilised for biomarker characterisation and validation. However there are caveats to their use, namely antigen preservation. The use of multicolour automated protocols, which utilise a house keeping antigen, circumvents these caveats and releases the full potential of TMAs in biomarker research.

328 Background: Outcome for many patients with high-grade muscle invasive bladder cancer (MIBC) remains poor despite aggressive therapy. This study used novel marker approaches to evaluate putative treatment targets, correlating cell line models of bladder cancer with a large scale MIBC tissue microarray (TMA) linked to long-term post-treatment outcomes. Methods: The TMA was constructed from 497 cystectomy specimens from patients with comprehensively documented disease outcome (minimum 5 years follow-up). TMA sections were stained for putative targets using DAB-based IHC. Cellular expression of CYR61 and CTGF was quantified (after targets had been identified using the genetic search programme, Oncomine) using Definiens automated image-analysis (AIA) software. Putative TMA based-targets studied were also evaluated in J82 and T24 bladder cancer cell-lines by siRNA knockdown during proliferation, time course stimulation and Boyden chamber MatrigelÒ invasion assays. Results: CTGF was increased significantly in both cell lines following stimulation with the cMET ligand HGF. siRNA knockdown of CTGF and CYR61 reduced cell proliferation and invasion towards HGF and FCS significantly (p

Prostate cancers ability to invade and grow in bone marrow stroma is thought to be due in part to degradative enzymes. The formation of prostate skeletal metastases have been reproduced in vitro by growing co-cultures of prostatic epithelial cells in bone marrow stroma. Expression of urokinase plasminogen activator, matrix metalloproteinase 1 and 7 by prostatic epithelial cells were identified using immunocytochemistry. Also, in vivo tissue sections from human prostatic bone marrow metastases were stained. To establish the role of these enzymes on colony formation, inhibitory antibodies directed against urokinase plasminogen activator, matrix metalloproteinase 1 and matrix metalloproteinase 7 were added into primary prostatic epithelial cells and bone marrow stroma co-cultures. All prostatic epithelial cell cultures stained positively for matrix metalloproteinase 1, matrix metalloproteinase 7 and urokinase plasminogen activator. Generally prostatic epithelial cells derived from malignant tissues showed increased staining in comparison to epithelia derived from non-malignant tissue. In agreement with in vitro co-cultures, the in vivo tissue sections of prostate bone marrow metastases showed positive staining for all three enzymes. Inhibition studies demonstrated that blocking matrix metalloproteinase 1, matrix metalloproteinase 7 and urokinase plasminogen activator function reduced the median epithelial colony area significantly in bone marrow stroma co-cultures in vitro. Using a human ex-vivo model we have shown that matrix metalloproteinase 1, matrix metalloproteinase 7 and urokinase plasminogen activator play an important role in the establishment of prostatic epithelial cells within bone marrow. British Journal of Cancer (2002) 86, 1136–1142. DOI: 10.1038/sj/bjc/6600207 www.bjcancer.com © 2002 Cancer Research UK

4744 Background: Prostate cancer is the most common malignancy in men with a predilection to metastasize to the bone. There is a requirement for a chemotherapeutic agent capable of preventing or sl...

Dissemination of tumour cells to the bone marrow is an early event in breast cancer, however cells may lie dormant for many years before bone metastases develop. Treatment for bone metastases is not curative, therefore new adjuvant therapies which prevent the colonisation of disseminated cells into metastatic lesions are required. There is evidence that cancer stem cells (CSCs) within breast tumours are capable of metastasis, but the mechanism by which these colonise bone is unknown. Here, we establish that bone marrow-derived IL1β stimulates breast cancer cell colonisation in the bone by inducing intracellular NFkB and CREB signalling in breast cancer cells, leading to autocrine Wnt signalling and CSC colony formation. Importantly, we show that inhibition of this pathway prevents both CSC colony formation in the bone environment, and bone metastasis. These findings establish that targeting IL1β-NFKB/CREB-Wnt signalling should be considered for adjuvant therapy to prevent breast cancer bone metastasis., In breast cancer, dormant cancer cells may develop into bone metastases. Here, the authors demonstrate that microenvironmental IL1β stimulates metastatic breast cancer cell colonisation in the bone via IL1β-NFKB/CREB-Wnt pathway activation and cancer stem cell colony formation

At present. a prognosis for prostate cancer (CaP) is determined by its accurate assessment of disease grade and stage. Histopathological typing using the Gleason grading system is the most universally accepted approach for grading CaP and provides an indication as to the aggressiveness of the tumour at the time of presentation. However, this system is based upon a visual criterion of pattern recognition that is operator dependent and subject to intra- and inter-observer variability, which can result in inappropriate patient management. Thus, there is a need for a molecular based diagnostic technique to grade tissue samples in a reliable and reproducible manner. In this paper we report a prototype diagnostic classifier for Gleason graded CaP tissue, based upon the integration of FTIR microspectroscopy with linear discriminant analysis (LDA). Blind testing of this model demonstrates 80% agreement of FTIR-LDA grade to histology, for the specimens analysed. We also study the effects of connective tissue absorption upon the area ratio of peaks at A1030 cm(-1)/A1080(cm(-1) which we use as a criterion to biospectroscopically map and distinguish areas of benign from malignant tissue. In addition, imaging time-of-flight secondary ion mass spectrometry (ToF-SIMS) has been applied to study freeze-dried, freeze-fractured prostate cancer cells in vitro. Preliminary results demonstrate localisation of various species including K, Ca and Mg within the cytoplasm that are present at millimolar concentrations and vital to cell physiology. The soft ionisation technique employed also permits for molecular information to be obtained and this has been used to evaluate chemically, different fracture planes within the analysis area.