thromboplastin and Prostatic-Neoplasms

Prostasomes are membrane-bound secretory vesicles produced by prostatic epithelial cells. They are known to carry many proteins, including tissue factor, and have membranes unusually rich in cholesterol and sphingomyelin. Prostasomes have well-documented effects on fertility, promoting sperm motility, stabilizing the acrosome reaction, and facilitating immunosuppression. This article reviews the evidence of the effects of prostasomes on in vitro angiogenesis assays, and the mechanism by which these effects occur. Seminal prostasomes seem to inhibit angiogenesis, whereas the equivalent particles released by malignant prostate cells promote angiogenesis. In both cases, the effects seem preserved after heat treatment to denature the protein content, suggesting an important role for lipid transfer, in particular, transfer of sphingomyelin.

Topics: Acrosome Reaction; Biological Transport; Cholesterol; Epithelial Cells; Humans; Male; Neovascularization, Pathologic; Prostate; Prostatic Neoplasms; Secretory Vesicles; Sperm Motility; Spermatozoa; Sphingomyelins; Thromboplastin

Thromboembolism is well recognized as a major complication of cancer. Many tumor cells overexpress tissue factor (TF), which activates blood coagulation in cancer patients. Inflammatory cells expressing TF are also contributors to this activation. In prostate cancer, we believe that prostasomes may also be involved in the initiation of blood coagulation. Prostasomes are submicron secretory granules derived from the prostate gland. They are surrounded by membrane and their extracellular appearance and membrane architecture are complex. Seminal prostasomes are believed to be necessary for successful fertilization and act as protectors of the spermatozoa in the lower and upper female genital tract. Cells from prostate cancer and its metastases are able to produce and export prostasomes to the extracellular environment. These prostasomes may differ quantitatively rather than qualitatively from their normal counterparts with regard to protein composition and function. A majority of human prostate cancers have been found to overexpress TF, and we have demonstrated by various methods that prostasomes derived from prostate cancer cells express considerably higher levels of TF compared with prostasomes of nonmalignant cell origin. The mechanism related to thromboembolic disease generated by prostasomes in prostatic cancer patients may be the early release of prostasomes from prostate cancer cells into the blood circulation, where they will evoke their blood-clotting effects.

Topics: Female; Gene Expression Regulation, Neoplastic; Humans; Male; Prostate; Prostatic Neoplasms; Secretory Vesicles; Semen; Sperm Capacitation; Sperm Motility; Spermatozoa; Thromboembolism; Thromboplastin

Topics: Blood Transfusion; Enzyme Inhibitors; Female; Fibrin; Fibrinogen; Fibrinolysin; Fibrinolysis; Hemostatics; Humans; Leukemia; Liver Cirrhosis; Male; Pathology; Physiology; Polycythemia Vera; Pregnancy; Pregnancy Complications; Pregnancy Complications, Hematologic; Prostatectomy; Prostatic Neoplasms; Prothrombin; Thrombin; Thromboplastin

To examine the hypothesis that increased monocyte tissue factor (mTF) levels may reflect urological tumour presence and progression.. Using a two-stage kinetic chromogenic assay, mTF levels were measured in 60 controls (normal subjects [60] and patients awaiting hernia repair or cholecystectomy [60]), patients with benign and malignant disease of the bladder (73), or prostate (81), and in patients with and without recurrent malignant disease of the bladder (30). The levels were assessed under fresh resting conditions (baseline) and after incubation for 6 h without (unstimulated) and with (stimulated) Escherichia coli endotoxin. Each benign disease group was subdivided into inflammatory and non-inflammatory categories.. Patients with bladder and prostate malignancy showed significantly higher mTF levels than did each control for baseline and stimulated cells. The benign inflammatory groups for both organs had significantly higher mTF levels than had each control for baseline cells. There was no difference between malignant and benign inflammatory groups. Stimulated mTF levels showed better discrimination between the study groups. The mTF levels were associated with histological tumour progression, serum prostate specific antigen level and static bone scan images. Levels were also higher in patients with bladder cancer recurrence than in those with a normal check cystoscopy.. Stimulated mTF levels are raised in malignant and inflammatory disease compared with controls and patients with non-inflammatory conditions, and give maximal discrimination between these groups. mTF levels showed an association with tumour grade and other markers of tumour progression.

Topics: Biomarkers, Tumor; Disease Progression; Female; Humans; Male; Monocytes; Neoplasm Recurrence, Local; Prognosis; Prostatic Neoplasms; Thromboplastin; Urinary Bladder Neoplasms

Increased risk of thromboembolic events is associated with prostate cancer, specifically linked to activation of tissue factor. Vitamin D has potential anticoagulant effects by the downregulation of tissue factor expression.. To evaluate the effects on clot formation, the morphological and viscoelastic profiles of prostate cancer patients, before and after ex vivo supplementation of Vitamin D was studied.. Participants were recruited into a metastatic, non-metastatic and reference group. Whole blood samples were treated ex vivo with a dose of 0.5μg/kg Calcitriol. Clot kinetics were assessed using Thromboelastography®. Morphology of the blood components were studied using scanning electron microscopy (SEM).. Results from the Thromboelastography® and SEM indicated no major differences between the non-metastatic group before and after treatment compared to the reference group. The Thromboelastography® showed that the metastatic group had an increased viscoelastic profile relating to a hypercoagulable state. Visible changes with regards to platelet activation and fibrin morphology were demonstrated with SEM analysis of the metastatic group. The viscoelastic and morphological properties for the non-metastatic group after treatment improved to be comparable to the reference group.. Vitamin D supplementation may lead to a more favorable viscoelastic profile, with less dangerous clots forming.

Topics: Dietary Supplements; Fibrin; Humans; Male; Prostatic Neoplasms; Thrombelastography; Thromboplastin; Thrombosis; Vitamin D

Essentials Androgen deprivation increases the rate of venous thromboembolism in prostate cancer patients. We characterized androgen receptor-mediated tissue factor regulation in prostate epithelial cells. Androgen receptor is dampening tissue factor expression in prostate epithelial cells. Androgen deprivation could enhance tissue factor expression and raise venous thromboembolism rates.. Background Prostate cancer is one of the leading causes of cancer death in men. Advanced prostate cancer is usually treated by androgen deprivation therapy (ADT), which is aimed at reducing circulating testosterone levels to reduce cancer growth. There is growing evidence that ADT can increase the rate of venous thromboembolism (VTE) in prostate cancer patients. The tissue factor (TF) gene is one of the most important mediators of coagulation and VTE, but, so far, there are limited data on androgen receptor (AR)-mediated TF gene expression. Objectives To characterize AR-mediated TF regulation in vitro and in vivo. Methods We used the androgen-dependent prostate cancer cell lines LNCaP and MyC-CaP to test whether TF expression is regulated by AR. Furthermore, we cloned the TF gene promoter into a luciferase reporter vector to identify the transcription factor-binding sites that mediate TF regulation downstream of AR. Finally, we used castration experiments in mice to characterize AR-mediated TF regulation in vivo. Results TF is directly regulated by AR. In LNCaP cells, nuclear factor-κB signaling and EGR1 mediate TF expression. By using castration experiments in mice, we could detect upregulation of TF and early growth response protein 1 mRNA and protein expression in prostate epithelial cells. Conclusion AR is crucial for dampening TF expression, which could be important for increased TF expression and TF-positive microvesicle release in androgen-deprived prostate cancer patients.

Topics: Androgen Antagonists; Androgens; Animals; Binding Sites; Cell Line, Tumor; Dihydrotestosterone; Down-Regulation; Early Growth Response Protein 1; Epithelial Cells; Humans; Male; Mice, Inbred C57BL; NF-kappa B; Orchiectomy; Promoter Regions, Genetic; Prostate; Prostatic Neoplasms; Protein Binding; Receptors, Androgen; Signal Transduction; Thromboplastin; Venous Thromboembolism

Prostate cancer (PC) patients in advanced stages of the disease have high risk of blood coagulation complications. The procoagulant molecule Tissue factor (TF), and the fibrinolysis inhibitor plasminogen activator inhibitor-1 PAI-1 play important role in this complication. Extracellular vesicles (EV) shed from cancer cells may contribute to the regulation of TF and PAI-1. The procoagulant activity of EV can be associated with the oncogenic and metastatic characteristics of their cells.. We have expressed EGFRvIII in DU145 cells to assess the role of this oncogene in the procoagulant activity of EV. The intercellular exchange of TF via EV was assessed by downregulating its expression in DU145 cells using shRNA vector, and determining the transfer of TF via EV enriched with the protein. Two PC cell lines with different metastatic potential were used to assess the correlation between the procoagulant activity of EV and the metastatic potential of PC cells. Photometric assays were used to determine FXa-activity and thrombin generation as indicators for the procoagulant activity of EV. Double-tagged proteinase-activated receptor 1(PAR-1) expressed in CHO cells to assess its activation by EV.. The expression of EGFRvIII in DU145 cells led to increased mRNA levels for TF and PAI-1, but the increase in these proteins expression was detected mostly in the EV. EV with enhanced levels of TF protein conferred higher TF procoagulant activity on the acceptor cells by intercellular exchange of this protein. Procoagulant activity of EV, assessed by FXa activity, and thrombin generation, was correlated with the oncogenic and metastatic potential of PC cells. The ability of EV to generate thrombin led to the activation of PAR-1, which was evident by the truncation of tagged-PAR-1.. The active oncogene EGFRvIII increases the concentration of TF and PAI-1 in EV. The procoagulant activity of EV is associated with the oncogenic and metastatic characteristics of their PC cells. Also, EV may contribute to the high procoagulant activity in the tumour microenvironment by the intercellular exchange of TF. Finally, through the generation of thrombin, EV can activate PAR-1, which evidently contributes to cancer progression, linking the coagulation system to tumor progression.

Topics: Cell Line, Tumor; Extracellular Vesicles; Humans; Male; Neoplasm Metastasis; Plasminogen Activator Inhibitor 1; Prostatic Neoplasms; Thrombin; Thromboplastin

Topics: Biomarkers; Humans; Prostatic Neoplasms; Thromboplastin; Thrombosis

Tumor cells display on their surface several molecular chaperones that normally reside in the endoplasmic reticulum. Because this display is unique to cancer cells, these chaperones are attractive targets for drug development. Previous epitope-mapping of autoantibodies (AutoAbs) from prostate cancer patients identified the 78-kDa glucose-regulated protein (GRP78) as one such target. Although we previously showed that anti-GRP78 AutoAbs increase tissue factor (TF) procoagulant activity on the surface of tumor cells, the direct effect of TF activation on tumor growth was not examined. In this study, we explore the interplay between the AutoAbs against cell surface-associated GRP78, TF expression/activity, and prostate cancer progression. First, we show that tumor GRP78 expression correlates with disease stage and that anti-GRP78 AutoAb levels parallel prostate-specific antigen concentrations in patient-derived serum samples. Second, we demonstrate that these anti-GRP78 AutoAbs target cell-surface GRP78, activating the unfolded protein response and inducing tumor cell proliferation through a TF-dependent mechanism, a specific effect reversed by neutralization or immunodepletion of the AutoAb pool. Finally, these AutoAbs enhance tumor growth in mice bearing human prostate cancer xenografts, and heparin derivatives specifically abrogate this effect by blocking AutoAb binding to cell-surface GRP78 and decreasing TF expression/activity. Together, these results establish a molecular mechanism in which AutoAbs against cell-surface GRP78 drive TF-mediated tumor progression in an experimental model of prostate cancer. Heparin derivatives counteract this mechanism and, as such, represent potentially appealing compounds to be evaluated in well-designed translational clinical trials.

Topics: Animals; Antineoplastic Agents; Autoantibodies; Cell Line, Tumor; Cell Membrane; Cell Proliferation; Endoplasmic Reticulum Chaperone BiP; Heat-Shock Proteins; Humans; Male; Mice, Inbred NOD; Mice, SCID; Neoplasm Grading; Neoplasm Proteins; Neoplasm Staging; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Random Allocation; Recombinant Proteins; Surface Properties; Thromboplastin; Tumor Burden; Unfolded Protein Response; Xenograft Model Antitumor Assays

A previously reported expression signature of three genes (IGFBP3, F3 and VGLL3) was shown to have potential prognostic value in estimating overall and cancer-specific survivals at diagnosis of prostate cancer in a pilot cohort study using freshly frozen Fine Needle Aspiration (FNA) samples.. We carried out a new cohort study with 241 prostate cancer patients diagnosed from 2004-2007 with a follow-up exceeding 6 years in order to verify the prognostic value of gene expression signature in formalin fixed paraffin embedded (FFPE) prostate core needle biopsy tissue samples. The cohort consisted of four patient groups with different survival times and death causes. A four multiplex one-step RT-qPCR test kit, designed and optimized for measuring the expression signature in FFPE core needle biopsy samples, was used. In archive FFPE biopsy samples the expression differences of two genes (IGFBP3 and F3) were measured. The survival time predictions using the current clinical parameters only, such as age at diagnosis, Gleason score, PSA value and tumor stage, and clinical parameters supplemented with the expression levels of IGFBP3 and F3, were compared.. When combined with currently used clinical parameters, the gene expression levels of IGFBP3 and F3 are improving the prediction of survival time as compared to using clinical parameters alone.. The assessment of IGFBP3 and F3 gene expression levels in FFPE prostate cancer tissue would provide an improved survival prediction for prostate cancer patients at the time of diagnosis.

Topics: Aged; Aged, 80 and over; Biopsy, Large-Core Needle; Cohort Studies; Formaldehyde; Gene Expression Regulation, Neoplastic; Humans; Insulin-Like Growth Factor Binding Protein 3; Linear Models; Logistic Models; Male; Middle Aged; Neoplasm Grading; Neoplasm Staging; Paraffin Embedding; Prognosis; Prostate; Prostatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; Survival Analysis; Thromboplastin; Tissue Fixation

Contrast-enhanced ultrasound with targeted microbubble contrast agents is an emerging technique for imaging biological processes at the molecular level. The accumulation of targeted microbubbles at tissue sites overexpressing specific molecular markers increases the backscattered signal for noninvasive evaluations of diseases. The aim of this preliminary study was to combine molecular imaging with an in vivo contrast agent quantification to support the early diagnosis of the pathology and to enhance the assessment of neoplastic tissues. Tumor growth was induced by subcutaneous injection of prostate cancer cells in four rats. Microbubbles targeted to tissue factor (TF) were administered. A vascularized region located in proximity to the tumor and centered around the focus depth was analyzed in each animal. The backscattered signals (i.e. the radio-frequency data) were acquired during two different perfusion conditions to evaluate the contribution of attached microbubbles. After image generation by means of a multi-pulse contrast-enhanced technique, a nonlinear regression method based on the support vector machine was employed to estimate the contrast agent concentrations in cubic voxels (1-mm side length). The number of attached microbubbles per mm(3) was estimated based on a multi-dimensional vector of features extracted from the processed radio-frequency signals. A significant correlation (p < 0.05) between the size of the tumors and the estimated microbubble concentration was found, thus opening the possibility for combining molecular imaging and contrast agent concentration mapping to refine pathology evaluation. Copyright © 2016 John Wiley & Sons, Ltd.

Topics: Animals; Contrast Media; Isoantibodies; Male; Microbubbles; Molecular Imaging; Prostatic Neoplasms; Rats; Thromboplastin; Ultrasonography

The aim of the study was to further investigate the role of fibrinogen-like protein 2 (FGL-2), a transmembrane prothrombinase that directly cleaves prothrombin to thrombin, in angiogenesis and tumor development and the mechanism(s) underlying these processes. To study angiogenesis HUVEC clones with decreased fgl-2 mRNA were generated by specific siRNA. To study tumorigenesis SCID mice were implanted with intact (wild type) and fgl-2-silenced PC-3 clones. IFN-γ treated HUVEC expressing increased fgl-2 mRNA exhibited significant capillary sprouting that was not inhibited by hirudin, whereas fgl-2 silencing completely inhibited blood-vessel formation. Tumors (poorly differentiated carcinoma) developed in all 12 mice injected with wild type PC-3 compared with 8/12 mice injected with the fgl-2-silenced PC-3 clone. The tumors developed by fgl-2-silenced PC-3 clones were smaller and less aggressive and contained significantly fewer blood vessels (p<0.05). All tumors' sections were negative for thrombin staining, indicating that FGL-2-induced tumorigenesis was not mediated by thrombin. In fgl-2-silenced tumors there was a decrease in fgl-2 mRNA (p=0.02) and ERK1/2 phosphorylation (p<0.05) by 80% and a 20%, respectively. The mechanism underlying these processes, studied in PC-3 clones, revealed that fgl-2 silencing was associated with a 65% decrease in FGF-2 mRNA (p<0.01) and a 30% down regulation of ERK1/2 phosphorylation (p<0.05). Together, these results suggest that FGL-2 mediates angiogenesis and tumorigenesis not by thrombin-mediated mechanism but rather through FGF-2/ERK signaling pathway. FGL-2 may serve as a valuable therapeutic target in the future.

Topics: Animals; Carcinogenesis; Cell Line, Tumor; Cell Proliferation; Fibrinogen; Human Umbilical Vein Endothelial Cells; Humans; Male; MAP Kinase Signaling System; Mice, SCID; Neovascularization, Pathologic; Prostate; Prostatic Neoplasms; RNA Interference; RNA, Small Interfering; Thrombin; Thromboplastin

Topics: Animals; Carcinogenesis; Fibrinogen; Humans; Male; MAP Kinase Signaling System; Neovascularization, Pathologic; Prostatic Neoplasms; Thromboplastin

The association between cancer and thrombogenesis has been recognized since 1865, and tissue factor (TF) is important at various stages in the natural history of the disease. It is involved in cancer angiogenesis, growth and metastasis. TF pathway inhibitor (TFPI), being the major physiological regulator of the TF-dependent coagulation pathway, is also important in establishing net procoagulant potential. In this study, we determine TF and TFPI levels in three prostate epithelial cell lines, one of normal and two of malignant origin. Cells were grown in standard maintenance conditions and harvested at more than 90% confluence. These were fractionated into cytosol, membrane and nuclei for analysis. Microparticles secreted into the culture medium were also analysed. TF and TFPI levels were determined using an ELISA. TF expression in these cells was also visualized using immunocytochemistry. There was absence of TF and TFPI in nuclei of all cell lines. TF expression was higher in subcellular fractions and microparticles of normal prostate cells than cancer cells. In contrast, levels of TFPI (structurally resembling a secreted, rather than transmembrane protein) in microparticles of normal prostate cells were much lower than tumour cells. In conclusion, the activity of prostate cancer cells themselves is unlikely to be the source of hypercoagulability in patients, but might precipitate chains of events that would produce such an effect.

Topics: Cell Line, Tumor; Cell Membrane; Cell Nucleus; Cell-Derived Microparticles; Cytosol; Gene Expression; Humans; Lipoproteins; Male; Prostate; Prostatic Neoplasms; Thrombophilia; Thromboplastin; Thrombosis

This study determines the impact of tissue factor (TF)-signaling on the extrinsic pathway of apoptosis in cancer cells and propose death associated protein kinase-1 (DAPK1) as a novel key regulator.. In MDA-MB-231 breast and PC3 prostate cancer cells, mRNA levels were analyzed by real-time PCR and protein expressions were assessed by flow cytometry or western blot. Caspase-8 and -3 levels, cell size, and changes in nuclear morphology were recorded using the ArrayScan microscope and 84 apoptosis-related genes were screened with the RT2 Profiler™ PCR Array.. In serum starved MDA-MB-231 cells, a TF/FVIIa-sensitive upregulation of apoptosis markers was recorded. Similarly, TRAIL-induced apoptosis was negatively regulated by TF/FVIIa (10 and 100 nM) and TF/FVIIa/FXa but not by active-site inhibited FVIIa. FVIIa, moreover, decreased the transcription of DAPK1 and thereby diminished the association between DAPK1 and FADD in the caspase-8 activating death-inducing signaling complex (DISC). TF/FVIIa regulation of caspase-8 and DAPK1 was dependent on PI3-kinase/AKT and independent of the protease activated receptors (PAR) 1 and 2. Despite of receptor expression and functional signaling, both PAR-agonist treatment and PAR-blocking antibodies in combination with FVIIa failed to influence the anti-apoptotic signal.. We hereby report that TF/FVIIa-induced signaling governs the extrinsic pathway of apoptosis by reducing the levels of DAPK1 in the DISC independently of PAR1 and PAR2. This implies the conceivable involvement of cell surface components other than the PARs and entails the search for TF-dependent regulators of DAPK1 transcription.

Topics: Apoptosis; Apoptosis Regulatory Proteins; Breast Neoplasms; Calcium-Calmodulin-Dependent Protein Kinases; Caspase 8; Caspase Inhibitors; Cell Line, Tumor; Death-Associated Protein Kinases; Enzyme Activation; Factor VIIa; Factor Xa; Fas-Associated Death Domain Protein; Female; Humans; Male; Phosphatidylinositol 3-Kinases; Phosphorylation; Prostatic Neoplasms; Proto-Oncogene Proteins c-akt; Receptor, PAR-1; Receptor, PAR-2; Recombinant Proteins; RNA, Messenger; Signal Transduction; Thromboplastin

We investigated whether serum markers of angiogenesis endothelin-1 (ET-1) and tissue factor (TF), and/or markers of vascular damage such as circulating endothelial cells (CECs), or their relative changes during treatment, were prognostic for overall survival (OS) in castration resistant prostate cancer (CRPC) patients. Additionally, we combined these markers with circulating tumour cells (CTCs) to construct a predictive nomogram for treatment outcome.. One hundred and sixty two CRPC patients treated with a docetaxel containing regimen had blood drawn before and at 2-5 weeks and 6-8 weeks after treatment start. Prospectively determined CTC and CEC levels, and retrospectively measured serum concentrations of ET-1 (pg/mL) and TF (pg/mL) were evaluated to determine their prognostic value for OS.. Baseline CEC, TF and ET-1 were not prognostic for OS. A > or = 3.8-fold increase in CEC 2-5 weeks after treatment initiation was associated with decreased OS (median 10.9 versus 16.8 months; P=0.015), as was any decrease in TF levels compared to baseline levels (median 11.9 versus 21.5 months; P=0.0005). As previously published, baseline and CTC counts > or = 5 at 2-5 weeks were also predictive of decreased OS. Combining CTC with changes in TF and CEC 2-5 weeks after treatment initiation yielded four groups differing in OS (median OS 24.2 versus 16.0 versus 11.4 versus 6.1 months; P<0.0001).. CEC, CTC and TF levels alone and combined can predict early on OS in CRPC patients treated with docetaxel-based therapy. A prospective study to confirm the use of these markers for patient management is needed.

Topics: Aged; Aged, 80 and over; Androgen Antagonists; Antineoplastic Agents; Biomarkers, Tumor; Docetaxel; Endothelin-1; Enzyme-Linked Immunosorbent Assay; Humans; Male; Middle Aged; Neoplastic Cells, Circulating; Neovascularization, Pathologic; Orchiectomy; Prospective Studies; Prostatic Neoplasms; Survival Analysis; Taxoids; Thromboplastin; Treatment Outcome

The increased risk of venous thromboembolism in cancer patients has been attributed to enhanced tissue factor (TF) procoagulant activity (PCA) on the surface of cancer cells. Recent studies have shown that TF PCA can be modulated by GRP78, an endoplasmic reticulum (ER)-resident molecular chaperone. In this study, we investigated the role of cell surface GRP78 in modulating TF PCA in several human cancer cell lines. Although both GRP78 and TF are present on the cell surface of cancer cells, there was no evidence of a stable interaction between recombinant human GRP78 and TF, nor was there any effect of exogenously added recombinant GRP78 on cell surface TF PCA. Treatment of cells with the ER stress-inducing agent thapsigargin, an inhibitor of the sarco(endo)plasmic reticulum Ca(2+) pump that causes Ca(2+) efflux from ER stores, increased cytosolic [Ca(2+)] and induced TF PCA. Consistent with these findings, anti-GRP78 autoantibodies that were isolated from the serum of patients with prostate cancer and bind to a specific N-terminal epitope (Leu(98)-Leu(115)) on cell surface GRP78, caused a dose-dependent increase in cytosolic [Ca(2+)] and enhanced TF PCA. The ability to interfere with cell surface GRP78 binding, block phospholipase C activity, sequester ER Ca(2+), or prevent plasma membrane phosphatidylserine exposure resulted in a significant decrease in the TF PCA induced by anti-GRP78 autoantibodies. Taken together, these findings provide evidence that engagement of the anti-GRP78 autoantibodies with cell surface GRP78 increases TF PCA through a mechanism that involves the release of Ca(2+) from ER stores. Furthermore, blocking GRP78 signaling on the surface of cancer cells attenuates TF PCA and has the potential to reduce the risk of cancer-related venous thromboembolism.

Topics: Antibodies, Neoplasm; Autoantibodies; Calcium; Cell Line, Tumor; Endoplasmic Reticulum; Endoplasmic Reticulum Chaperone BiP; Enzyme Inhibitors; Epitopes; Heat-Shock Proteins; Humans; Male; Phosphatidylserines; Prostatic Neoplasms; Sarcoplasmic Reticulum Calcium-Transporting ATPases; Signal Transduction; Thapsigargin; Thromboplastin; Type C Phospholipases; Venous Thromboembolism

Tissue factor (TF) is a cell surface glycoprotein intricately related to blood coagulation and inflammation. This study was performed to investigate the role of monocyte-lineage cells in prostate cancer cell TF expression and cell invasion.. Prostate cancer cell invasion was tested with and without added peripheral blood monocytes or human monocyte-lineage cell lines. TF neutralizing antibodies were used to determine the TF requirement for prostate cancer cell invasion activity. Immunohistochemistry was performed to identify prostate tissue CD68 positive monocyte-derived cells and prostate epithelial TF expression.. Co-culture of PC-3, DU145, and LNCaP cells with isolated human monocytes significantly stimulated prostate cancer cell invasion activity. TF expression was greater in highly invasive prostate cancer cells and was induced in PC-3, DU145, and LNCaP cells by co-culture with U-937 cells, but not with THP-1 cells. TF neutralizing antibodies inhibited PC-3 cell invasion in co-cultures with monocyte-lineage U-937 or THP-1 cells. Prostate cancer tissues contained more CD68 positive cells in the stroma and epithelium (145 ± 53/mm(2)) than benign prostate (108 ± 31/mm(2)). Samples from advanced stage prostate cancer tended to contain more CD68 positive cells when compared with lower stage lesions. Prostatic adenocarcinoma demonstrated significantly increased TF expression compared with benign prostatic epithelium.. This study shows that co-culture with monocyte-lineage cells induced prostate cancer cell invasion activity. PC-3 invasion and TF expression was induced in co-culture with U-937 cells and partially inhibited with TF neutralizing antibodies.

Topics: Adenocarcinoma; Cell Growth Processes; Cell Line, Tumor; Cell Movement; Coculture Techniques; Humans; Immunohistochemistry; Male; Monocytes; Prostatic Neoplasms; Receptor, Anaphylatoxin C5a; Receptors, Complement; Statistics, Nonparametric; Thromboplastin; Tissue Array Analysis

Tissue factor (TF) is the principal physiologic initiator of coagulation. It also plays an important role in tumor growth and metastasis possibly by contributing to angiogenesis. We evaluated the expression of TF in benign and malignant prostate tissue and correlated it with the expression of the pro-angiogenic protein, vascular endothelial growth factor (VEGF). We used a tissue microarray (TMA) constructed from 80 archival prostatectomy specimens. Core samples were collected from benign prostate tissue (BP) (n= 77), high-grade prostatic intraepithelial neoplasia (PIN) (n= 26), and carcinoma (PCa) (n= 93). TMA sections were stained with an immunopurified polyclonal TF antibody and a rabbit polyclonal VEGF. Two pathologists manually scored staining in epithelial cells using the German Immunoreactive Score. Positive staining for TF was seen predominantly in PCa with rare positive glands in BP and PIN. TF expression was significantly lower in BP versus PCa specimens (p< .001) and in PIN versus PCa specimens (p< .001). Positive staining for VEGF was seen in PCa, BP, and PIN. Rates of VEGF expression were also significantly lower in BP versus PCa specimens (p= .003) but not in PCa versus PIN (p= .430). The majority of PCa samples positive for TF were also positive for VEGF (p< .001). Our findings reinforce the link between angiogenesis and TF expression in PCa. We suggest further exploration of TF-mediated pathways leading to increased tumor aggressiveness in PCa, and the possible use of anti-TF agents in PCa.

Topics: Carcinoma; Humans; Immunohistochemistry; Male; Neovascularization, Pathologic; Prostatectomy; Prostatic Intraepithelial Neoplasia; Prostatic Neoplasms; Thromboplastin; Tissue Array Analysis; Vascular Endothelial Growth Factor A

Tissue factor (TF) plays a critical role in tumour growth and metastasis, and its enhanced release into plasma in association with cellular microparticles (MPs) has recently been associated with pathological cancer progression. We have previously demonstrated significantly elevated levels of plasma TF antigen as well as systemic coagulation and platelet activation in patients with localised prostate cancer. In this prospective study, we used a highly sensitive one-stage clotting assay to measure preoperative TF-specific procoagulant activity (PCA) of plasma MPs in 68 consecutive patients with early-stage prostate cancer to further explore the relevance of circulating TF in this tumour entity. Automated calibrated thrombography was used to monitor thrombin generation in cell-free plasma samples in the absence of exogenous TF or phospholipids. Compared to healthy male controls (n=20), patients had significantly increased levels of both D-dimer and TF-specific PCA of plasma MPs (p<0.001). Furthermore, MP-associated TF PCA was higher in patients with (n=29) than in those without (n=39) laboratory evidence of an acute-phase reaction (p=0.004) and decreased to normal levels within one week after radical prostatectomy. Overall, we found a significant correlation between TF-specific PCA of plasma MPs and plasma D-dimer (p=0.002), suggesting that plasma MPs contributed to in-vivo coagulation activation in a TF-dependent manner. Thrombin generation in plasma was also significantly increased in patients compared to controls (p<0.01). Collectively, our findings suggest that TF-specific PCA of plasma MPs contributes to intravascular coagulation activation in patients with early-stage prostate cancer and may represent a potential link between hypercoagulability, inflammation, and disease progression.

Topics: Acute-Phase Reaction; Aged; Antibodies, Monoclonal; Blood Coagulation; Blood Coagulation Tests; Cell Line, Tumor; Cell Separation; Cell-Derived Microparticles; Disease Progression; Flow Cytometry; Humans; Lipopolysaccharides; Male; Middle Aged; Neoplasm Staging; Prospective Studies; Prostatic Neoplasms; Sensitivity and Specificity; Thrombin; Thromboplastin

Cancer confers a prothrombotic state and statins are associated with a lowered risk for prostate cancer in vivo by unknown mechanisms. Prostate cancer cells release tissue factor (TF)-bearing, cholesterol-rich prostasomes which are pro-coagulant in vitro and a possible source for the blood-borne TF found in prostate cancer patients. We investigated the effect of cholesterol depletion on the production of prostasomes and on the TF activity in the conditioned medium of simvastatin-treated PC3 cells. Human PC3 prostate cancer cells were treated with high and low concentrations of simvastatin for different time periods. Caspase-3 was detected with the Array Scan microscope, whereas TF mRNA and protein were analyzed by TaqMan and flow cytometry. TF activity was assessed by measuring the cleavage of a chromogenic thrombin substrate. Prostasomes were isolated by repeated centrifugations and detected and quantified by flow cytometry. A micromolar dose of simvastatin caused reduction of TF expression and induction of apoptosis in the PC3 cells. The levels of TF on the prostasomes were also decreased but the TF activity in the conditioned medium of the simvastatin-treated PC3 cells was increased due to apoptosis-dependent release of prostasomes. Treatment with a nanomolar dose of simvastatin did not induce apoptosis or alter the expression of TF but instead decreased the production and release of the prostasomes. The TF activity was reduced in parity with the decline in prostasome release. In conclusion, in prostate cancer, a nanomolar dose of simvastatin may have an anti-thrombotic effect due to decreased levels of circulating TF-bearing prostasomes.

Topics: Anticholesteremic Agents; Apoptosis; Cell Line, Tumor; Culture Media, Conditioned; Dose-Response Relationship, Drug; Down-Regulation; Flow Cytometry; Humans; Male; Particle Size; Prostatic Neoplasms; Simvastatin; Thromboplastin; Venous Thrombosis

Tissue factor (TF), apart from its established role in hemostasis, has been implicated in promoting angiogenesis and metastasis in a wide array of tumors including prostate cancer. Expression of TF was evaluated in freshly-resected prostate specimens obtained from patients with localized (n=9) and androgen ablated (n=6) disease using real-time reverse transcription-polymerase chain reaction and Western blot analysis. TF was detected in all specimens in both stages of the disease. We further analyzed for correlations between TF expression and those of several angiogenic growth factors and their receptors. TF RNA expression correlated significantly with expression of vascular endothelial growth factor-A in these specimens (s=0.621, P=0.013). Eighty-one prostate specimens from patients with benign prostatic hyperplasia (n=27), localized prostate cancer (ES, n=32), and advanced disease (n=22) were also evaluated using immunohistochemistry and findings were correlated with clinical parameters. TF expression was detected on epithelial cells of the malignant glands. Furthermore, its expression levels correlated significantly with Gleason score (s=0.58, P=0.0001) and with the stage of the disease (s=0.441, P=0.0001) in these specimens. These data support the role of TF in angiogenesis and disease progression.

Topics: Adult; Aged; Aged, 80 and over; Disease Progression; Humans; Immunohistochemistry; Male; Middle Aged; Neoplasm Staging; Phenotype; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; RNA; Thromboplastin

Reducing expression of the tissue factor gene in prostate adenocarcinoma cells (PAIII) results in a cell line that, in vivo, mimics the growth of wildtype (wt) PAIII. However, instead of continuing to grow and metastasize as wt PAIII tumors do, tissue factor deficient PAIII (TFD PAIII) masses spontaneously regress after several weeks. Although whole cell vaccines are typically inactivated prior to administration to prevent proliferation within the host, numerous studies have suggested that exposure to live, attenuated, whole tumor cells, and the extracellular microenvironment they recruit, increases immunotherapeutic potential. Here, we provide support for this notion, and a strategy through which to implement it, by demonstrating that subcutaneous vaccinations with the TFD PAIII protect the Lobund-Wistar rat against subsequent wt PAIII cell challenge. TFD PAIII immunized rats suffered significantly less metastasis of wt PAIII challenge tumors compared to unvaccinated naïve controls rats. These results offer the intriguing possibility that the TFD PAIII vaccine is an effective system for the prevention and, possibly, the treatment of prostate cancer.

Topics: Adenocarcinoma; Animals; Cancer Vaccines; Cloning, Molecular; Immunotherapy; Male; Prostatic Neoplasms; Rats; Reverse Transcriptase Polymerase Chain Reaction; Thromboplastin; Transfection; Tumor Cells, Cultured

Prostasomes are secretory granules produced by the glandular epithelial cells of the prostate. Seminal prostasomes contain high amounts of Tissue Factor (TF) but no studies of TF on malignant cell prostasomes have been made. Here we compare the expression, phosphorylation, and function of TF on prostasomes of different origin.. TF was detected on prostasomes isolated from seminal fluid and human prostate cancer cell lines (PC-3, DU145, and LNCaP) using FACS and enzyme immunoassay (EIA). Incubation of prostasomes with radioactive ATP under conditions favoring protein kinase A activity led to phosphorylation of TF as detected by immunoprecipitation and SDS-PAGE. The prothrombotic effect of prostasomes was investigated in whole blood and recalcified plasma. Blocking experiments were performed using anti-TF antibodies and corn trypsin inhibitor.. TF was expressed on all tested prostasome preparations with lowest values found for seminal ones. Prostasomal TF was the main endogenous substrate for prostasomal protein kinase A. All tested prostasome preparations greatly enhanced the rate of clot formation in a dose-dependent fashion, that is, the clotting capability of prostasomes seemed to be related to the extent of their expression of TF. In addition, the density of the clot varied between different prostasome preparations. When incubated in whole blood, prostasomes were found to associate to WBC thereby inducing them to express and release TF.. These data show that TF is overexpressed and also subjected to phosphorylation by malignant cell prostasomes. This suggests major roles for prostasomes in thrombotic events that occur in some advanced cases of prostate cancer.

Topics: Calcium; Cell Line, Tumor; Flow Cytometry; Humans; Immunoenzyme Techniques; Male; Nephelometry and Turbidimetry; Phosphorylation; Prostate; Prostatic Neoplasms; Protein Kinases; Secretory Vesicles; Seminiferous Epithelium; Thromboplastin; Thrombosis

Topics: Biomarkers, Tumor; Blood Coagulation; Cysteine Endopeptidases; Humans; Male; Neoplasm Proteins; Prognosis; Prostatic Neoplasms; Thromboplastin

Tissue factor (TF) is involved in cancer growth and metastasis, and haemostatic abnormalities are found in most patients with advanced malignancies, including prostate cancer (PC). Because anti-haemostatic agents are increasingly screened for their potential to prolong survival in tumor patients, a detailed characterization of haemostatic markers in selected cancer subtypes and clinical stages is warranted. In this study, we measured preoperative plasma TF antigen in a large cohort of patients with localized PC and correlated its levels with markers of coagulation and platelet activation, prostate-specific antigen (PSA), and histopathological findings to explore its potential as a prognostic marker in this tumor entity. Out of 140 patients, 19% and 23% had plasma TF antigen levels of <40 pg/ml (low-TF) and >200 pg/ml (high-TF), respectively, which was substantially higher than in 42 healthy male controls. Patients also had low-grade systemic coagulation activation as evidenced by elevated D-dimer, F1 + 2, and PAP plasma levels. Furthermore, similar to sP-selectin and sCD40L antigen, flow cytometric analysis of platelet-derived microparticles in plasma revealed significantly increased numbers in high-TF as compared to low-TF patients and controls. Whereas elevated D-dimer was associated with larger and less differentiated tumors, preoperative plasma TF antigen levels (median [IQR]) were higher in patients with (161 pg/ml [100-236]) than in those without recurrent PC (105 pg/ml [52-182]), as indicated by a serum PSA of >0.1 ng/ml during ambulatory follow-up. In patients with localized PC, preoperative plasma TF antigen levels correlate with platelet activation in vivo and may indicate an increased risk for recurrent disease.

Topics: alpha-2-Antiplasmin; Antigens, Neoplasm; Biomarkers, Tumor; Blood Coagulation; CD40 Ligand; Cell Differentiation; Disseminated Intravascular Coagulation; Enzyme-Linked Immunosorbent Assay; Fibrin Fibrinogen Degradation Products; Fibrinolysin; Flow Cytometry; Follow-Up Studies; Hemostasis; Humans; Male; Middle Aged; Neoplasm Invasiveness; P-Selectin; Peptide Fragments; Platelet Activation; Prognosis; Prospective Studies; Prostate-Specific Antigen; Prostatectomy; Prostatic Neoplasms; Prothrombin; Recurrence; Risk Assessment; Thromboplastin; Time Factors

Mouse factor VII (mfVII), ligand of tissue factor (TF) which is frequently over-expressed during neovascularization activated by tumor growth, was fused to staphylococcus enterotoxin A (SEA) that mediates greater intensity of T-cell activation against tumor cells. The anti-tumor effects of the mfVII-SEA chimeric protein were evaluated.. Fusion of SEA and mfVII cDNA was constructed using adenovirus vector and produced in 293 packaging cell lines. The 293 cells containing the adenovirus were administered subcutaneously to mice. Fluorescence studies at the injection site and the liver were performed 3 days later. Mouse prostatic tumor RM-1 cells and mouse sarcoma MCA 205 H12 cell lines were then used in mice to create lung metastasis and subcutaneous tumor to carry out efficacy evaluation, respectively.. Adenovirus released from the injected 293 cells only infected the subcutaneous tissue at the injection site. The in vivo experiments in mice revealed that formation of lung metastasis was strongly inhibited by the mfVII-SEA (23 +/- 8) compared to the vacant vector control group (193 +/- 38) and PBS control group (211 +/- 42) (P < 0.01). The mfVII-SEA also strongly suppressed tumor growth at the subcutaneous injection site (342.6 +/- 107.1) mm(3) compared to that of vacant vector control (2244.3 +/- 350) mm(3) and SEA (1208.3 +/- 210) mm(3) by the 23rd day.. The chimeric protein mfVII-SEA significantly inhibits lung metastasis formation and local tumor growth.

Topics: Animals; Antigens, Bacterial; Antineoplastic Agents; Enterotoxins; Factor VII; Female; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; Prostatic Neoplasms; Recombinant Fusion Proteins; Staphylococcus; Thromboplastin

In health, haemostasis and angiogenesis are tightly regulated processes, but may become deregulated in cancer. Recent evidence suggests that platelet activation may link these processes as platelets can release angiogenic factors such as vascular endothelial growth factor (VEGF). Furthermore, inflammation has also been implicated in regulating both coagulation and angiogenesis, possibly by activating platelets directly and increasing, for example, plasma fibrinogen. We hypothesized relationships between plasma markers of the processes in two common forms of cancer. Plasma levels of VEGF (reflecting angiogenesis), soluble P-selectin, (marking platelet activation), tissue factor [TF], fibrinogen and fibrin D-dimer (coagulation markers), and serum levels of IL-6 (inflammation) were measured by ELISA in 30 patients with biopsy-proven breast cancer, 30 patients with biopsy-proven prostate cancer, and 30 age- and sex-matched controls for each group. Prostate specific antigen was also measured in the men. Release of VEGF from IL-6 stimulated platelets was assessed by ELISA. Plasma levels of IL-6 (P <0.02), VEGF, soluble P-selectin, fibrinogen, and fibrin D-dimer (all p <0.01) were significantly raised in breast cancer, whereas VEGF, soluble P-selectin, fibrin D-dimer (all p <0.01) and fibrinogen (p <0.05) were significantly raised in prostate cancer. Significant correlations were found between IL-6 and VEGF (p <0.01), and IL-6 and soluble P-selectin (p = 0.038) in breast cancer. Further experiments demonstrated an in vitro IL-6 induced dose-dependent release of VEGF from platelets. In conclusion, strong relationships between IL6 and VEGF, but not with coagulation or platelet markers, and release of VEGF from IL-6 stimulated platelets, suggest a role for inflammation and platelets in angiogenesis.

Topics: Aged; Blood Coagulation; Breast Neoplasms; Case-Control Studies; Female; Humans; In Vitro Techniques; Interleukin-6; Male; Middle Aged; Neovascularization, Pathologic; P-Selectin; Platelet Activation; Prostatic Neoplasms; Thromboplastin; Vascular Endothelial Growth Factor A

Human tissue factor (TF) is involved in tumor angiogenesis and metastasis. However, little is known about the distribution of TF in urological cancer. In this study we investigated the TF expression in tumor tissue and autologous non-malignant tissue as well as in serum of patients with renal cell carcinoma (RCC), bladder cancer, and prostate cancer (PCa). To study the distribution of TF in tumor tissue and in the surrounding non-malignant tissue, we measured TF protein by ELISA in tissue specimens obtained intraoperatively from 18 RCC, seven bladder cancer and six PCa patients. Differences in TF expression were found between tumor tissue and nonmalignant tissue for the three tumor types at the protein level (in the order RCC < bladder cancer < PCa). In all but one of the 18 RCC patients (94 %) higher TF protein level was observed in non-malignant tissue as compared to the tumor tissue. In addition, the relative TF mRNA expression analyzed by a quantitative RT-PCR assay in the same RCC tissue sample pairs was higher in 78% of non-malignant tissues in comparison to the tumor tissue specimens. Moreover, using enzyme linked immunosorbent assay the TF protein content was measured in serum samples of 66 patients with bladder cancer, 75 RCC patients and 157 PCa patients, and was compared with the TF serum level of 92 healthy volunteers. Whereas no differences were detected between normal volunteers and patients with PCa or RCC, patients with bladder cancer showed a significantly increased level of serum TF (P=0.0076). However, no causal association between TF levels in serum and TF content in tissue extracts for all three tumor types of urological tumors was found. Our results suggest that TF in non-malignant renal tissues was expressed at a higher level compared to the supposed de novo TF expression in RCC tissue specimens. This indicates a tumor-associated induction of TF expression in the TF-negative RCC progenitor cells. The increased serum TF levels are alike the reported higher urinary TF levels found in bladder cancer patients. The potential clinical relevance of this finding should be further elucidated.

Topics: Carcinoma, Renal Cell; Enzyme-Linked Immunosorbent Assay; Humans; Male; Polymerase Chain Reaction; Prostatic Neoplasms; RNA, Messenger; Thromboplastin; Tissue Distribution; Tumor Cells, Cultured; Urinary Bladder Neoplasms; Urologic Neoplasms

To assess immunohistochemically the pattern of tissue factor (TF) expression in patients with metastatic prostate cancer, because TF is aberrantly expressed in human cancer. TF is the primary initiator of the coagulation cascade.. Seventy-three patients with untreated metastatic prostate cancer who received hormonal therapy were included in the present study. Biopsy specimens were stained with anti-human TF antibody. We evaluated the histologic grade, extent of bony metastasis, clinical response to hormonal therapy, and patient prognosis.. TF was detected in 75.3% of the tumors of the patients with metastatic prostate cancer. TF expression showed no association with histologic grade, extent of bony metastasis, or clinical response to hormonal therapy. Patients with TF-positive tumors had a poorer cause-specific survival than those with TF-negative tumors. Multivariate analysis showed that TF expression, clinical response to hormonal therapy, and extent of bony metastasis were significant prognostic factors.. The TF content measured using immunohistochemical staining was a useful prognostic factor for patients with metastatic prostate cancer treated with androgen withdrawal therapy.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Antineoplastic Agents, Hormonal; Bone Neoplasms; Chlormadinone Acetate; Diethylstilbestrol; Gene Expression Regulation, Neoplastic; Humans; Life Tables; Male; Middle Aged; Neoplasm Proteins; Orchiectomy; Prognosis; Proportional Hazards Models; Prostatic Neoplasms; Survival Analysis; Thromboplastin

Tissue factor (TF) is primarily known for its function to initiate blood coagulation. The range of in vivo expression of TF is wide and requires a dynamic assay for monitoring. A general method for TF mRNA quantitation that is dynamic, sensitive and applicable to a variety of experimental systems or clinical situations is therefore desirable.. To develop a method for sensitive and dynamic quantitation of TF mRNA in human blood cells.. TF mRNA expression was analysed and evaluated in monocyte isolations, in whole blood (healthy volunteers and patients scheduled for percutaneous coronary intervention, PCI) and in a panel of human cell lines. RNA was extracted, reverse transcribed and subjected to real-time PCR amplification, according to the TaqMan technology. A TF plasmid was constructed as calibrator of the assay. Two housekeeping genes used as endogenous controls for cDNA quality and integrity were evaluated.. The assay was linear by seven orders of magnitude and detected down to 10(2) copies of the TF plasmid. The coefficient of variation was 4% intra-assay and 28% between the assays when using beta2MG as endogenous control. The beta-actin gene expression was induced by treatment with lipopolysaccharide (LPS) in blood leukocytes and could not be used as an endogenous control. However, beta2MG showed only minor variations upon treatment with LPS. The TF mRNA and antigen expression, measured in a Western blot, correlated well (R(2)=0.903) in a panel of 11 human cell lines.. We have established a method for sensitive and dynamic quantitation of TF mRNA in experimental systems and for clinical situations.

Topics: Base Sequence; Breast Neoplasms; Calibration; Cell Line, Tumor; Cells, Cultured; DNA Primers; DNA Probes; Female; Humans; Male; Plasmids; Polymerase Chain Reaction; Prostatic Neoplasms; RNA; RNA, Messenger; Sensitivity and Specificity; Thromboplastin

Prostate-specific membrane antigen (PSMA), a glutamyl preferring carboxypeptidase, is found in prostate and other carcinomas present on both tumor cells and associated microvascular lining cells. We find that the channel structures delineated by PSMA-expressing cells in human and rat prostate tumors are in functional continuity with the vasculature and thus form part of tumor microvasculature. The PSMA-positive cell-outlined channels are CD31 negative and mutually exclusive of CD31-positive cell-lined channels elsewhere in the tumor consistent with tumor cells adapted to a pseudoendothelial phenotype in vasculogenic mimicry. To assess the functional potential of such PSMA-lined microvasculature to selectively direct infarctive tumor therapy, we coupled the soluble extracellular domain of tissue factor to a PSMA catalytic site inhibitor to create a PSMA-directed selective tumor vascular thrombogen (STVT). This protein induced selective local in vivo infarctive necrosis of the rat Mat Lu prostate tumor when administered i.v. The combined administration of this STVT with low-dose doxorubicin produced a profound tumoricidal effect, resulting in complete eradication of some tumors. This is consistent with the therapeutic potential for a PSMA-directed STVT and expands the potential for selective infarctive ablation of tumors.

Topics: Animals; Antigens, Surface; Antineoplastic Combined Chemotherapy Protocols; Binding Sites; Carboxypeptidases; Catalysis; Dipeptides; Doxorubicin; Glutamate Carboxypeptidase II; Humans; Infarction; Liposomes; Male; Mice; Mice, Nude; Prostatic Neoplasms; Rats; Thromboplastin; Thrombosis; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Tissue factor (TF), the initiator of the extrinsic pathway of coagulation, binds plasminogen (Pg) with high affinity through an interaction between kringles 1-3 of Pg and the extracellular domain of TF. We investigated the binding of Pg type 1 (Pg 1) and Pg type 2 (Pg 2) to highly invasive, TF-expressing, 1-LN human prostate tumor cells and to TF isolated from 1-LN cell membranes. Pg 1, containing both N-linked and O-linked oligosaccharide chains, bound to isolated TF with high affinity, whereas Pg 2, containing only one O-linked oligosaccharide chain, did not bind to TF. Although Pg 1 and Pg 2 bind to 1-LN cells, only anti-TF antibodies inhibited the binding of Pg 1, suggesting that TF functions as the receptor for Pg 1 on 1-LN cells. Binding of Pg 1 to isolated TF was inhibited by 6-aminohexanoic acid and alpha-methylmannoside, suggesting that Pg 1 L-lysine binding sites and the biantennary, mannose-containing N-linked oligosaccharide chain are involved in this interaction. Binding of Pg 1 to 1-LN cells promoted activation by receptor-bound urinary-type Pg activator (u-PA) and initiated a Ca(++) signaling cascade. In previous studies we demonstrated that the Pg 2 O-linked carbohydrate chain is essential for its binding to CD26 on 1-LN cells. The current studies suggest that Pg oligosaccharide chains regulate the binding of Pg 1 and Pg 2 to separate receptors on the cell surface.

Topics: Binding Sites; Calcium Signaling; Glycosylation; Humans; Male; Matrix Metalloproteinase 9; Neoplasm Invasiveness; Oligosaccharides; Plasminogen; Prostatic Neoplasms; Protein Binding; Protein Processing, Post-Translational; Thromboplastin; Tumor Cells, Cultured; Urokinase-Type Plasminogen Activator

Topics: Adenoma; Biomarkers, Tumor; Humans; Intestinal Neoplasms; Intestine, Large; Male; Neoplasms; Prognosis; Prostatic Neoplasms; Thromboplastin

Coagulation activation is a recognized complication of cancer in which increased tissue factor (TF) is implicated. TF can be detected in urine (uTF). This study assesses uTF levels in benign and malignant urological disease and correlates the results with conventional markers of tumour progression.. Using a simple and reproducible kinetic chromogenic assay, we determined uTF levels in controls (normal volunteers (n = 57) and patients with renal stones (n = 30)), benign and malignant bladder (n = 75) or prostate (n = 106) disease and in patients with or without recurrent bladder cancer (n=30). Each benign disease group was stratified as inflammatory (cystitis or prostatitis) or non-inflammatory (negative cystoscopy following haematuria or benign prostatic hypertrophy).. The controls and the benign non-inflammatory results were indistinguishable. The malignant and inflammatory groups showed raised uTF levels over controls (P<0.001 bladder and P<0.01 prostate). The difference between malignant and benign inflammatory disease was only significant for the bladder group. uTF levels were significantly related to histological tumour grading, prostate serum specific antigen, static bone scan images and recurrence status.. uTF levels can distinguish, statistically but not without overlap, patients with malignancy from normal controls and benign non-inflammatory conditions. Discrimination between inflammatory and malignant disease has only been demonstrated in the bladder. uTF levels showed a significant association with markers of tumour progression or metastasis and may be useful in predicting bladder tumour recurrence.

Topics: Adult; Aged; Biomarkers, Tumor; Bone Neoplasms; Case-Control Studies; Cystitis; Disease Progression; Humans; Kidney Calculi; Male; Middle Aged; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms; Prostatitis; Thromboplastin; Urinary Bladder Neoplasms

In tumors, the switch to the angiogenic phenotype is thought to be controlled by a balance of positive and negative angiogenic factors. Tissue factor (TF) produced by tumor cells has been implicated in the regulation of this "angiogenic switch" through its ability to concurrently induce the expression of angiogenic molecules such as vascular endothelial cell growth factor (VEGF), while inhibiting the expression of anti-angiogenic molecules such as thrombospondin 2. We have examined TF expression and its relationship to angiogenesis and tumor progression in human prostate carcinomas. Most of the prostate carcinoma specimens examined (73%; n = 67) express high levels of TF. Immunohistochemical analysis localized TF expression to the epithelial cells of malignant glands. TF expression was significantly correlated with tumor angiogenesis as measured by the microvessel density (MVD). In addition, TF expression was correlated with the preoperative PSA level, a strong predictor of recurrence in prostate carcinomas. Our findings show that TF expression by the malignant glands in prostate cancer is common and suggest a role for this molecule in regulating prostate cancer progression and angiogenesis.

Topics: Adenocarcinoma; Disease Progression; Humans; Immunohistochemistry; Male; Microcirculation; Middle Aged; Neovascularization, Pathologic; Prostatic Neoplasms; Thromboplastin

The association between cancer and thromboembolic disease has been known for over a century. Increased tissue factor expression by endothelial cells, monocytes or macrophages is implicated. Thus, monocyte tissue factor measurements may reflect disease presence or progression.. Using a 2 stage kinetic chromogenic assay, monocyte tissue factor levels were assessed in normal controls (n=60), patient controls (hernia or cholecystectomy, n=60) and in patients with benign and malignant disease of the bladder (n=73), prostate (n=81), breast (n=83) and colorectum (n=62). This was performed as baseline (resting cells) and after 6 hours incubation with (stimulated) and without (unstimulated) lipopolysaccharide. Each benign disease group was sub-divided into inflammatory and non-inflammatory categories.. The relative operating characteristic curve for the lipopolysaccharide-stimulated monocyte tissue factor assay showed sensitivity and specificity for cancer, the area under the curve being 0.71. The control groups and the benign non-inflammatory groups gave similar results and were pooled for further analysis. Each malignant group showed higher monocyte tissue factor levels than the control groups for baseline (P< 0.05) and lipopolysaccharide-stimulated cells (P< 0.05). All benign inflammatory groups apart from breast, showed increased monocyte tissue factor levels over controls for baseline (P< 0.05) and lipopolysaccharide-stimulated cells (P< 0.05). In all cases there was no significant difference between the malignant and the benign inflammatory groups. Monocyte tissue factor levels were related to tumor grade or stage, patients' survival time, serum prostate specific antigen and static bone scan images. Levels were also higher in patients with bladder cancer recurrence and in those who subsequently died.. Lipopolysaccharide-stimulated monocyte tissue factor assay showed sensitivity and specificity for cancer compared to controls. Monocyte tissue factor levels are raised in malignant groups compared to controls and non-inflammatory diseases but not when compared with inflammatory conditions. Stimulated cells give better discrimination between the groups and may be useful in identifying high risk individuals. Monocyte tissue factor levels were related to tumor progression.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Breast Diseases; Breast Neoplasms; Case-Control Studies; Colonic Diseases; Colorectal Neoplasms; Discriminant Analysis; Disease Progression; Female; Humans; Inflammation; Male; Middle Aged; Monocytes; Prostatic Diseases; Prostatic Neoplasms; Risk Factors; Sensitivity and Specificity; Thromboembolism; Thromboplastin; Urinary Bladder Diseases; Urinary Bladder Neoplasms

Monocyte and urinary tissue factors (mTF and uTF) are both elevated in a number of pathologic conditions, including cancer. This study validates the best available uTF and mTF assays as diagnostic tools for cancer and examines if uTF levels reflect monocyte activation. Using kinetic chromogenic assays for uTF and mTF (measured on fresh resting cells [baseline], unstimulated cells, and lipopolysaccharide [LPS]-stimulated cells), we assessed TF levels in normal individuals, surgical controls, and patients with benign and malignant diseases. Each benign disease group was stratified as inflammatory or noninflammatory. Controls and benign noninflammatory results were indistinguishable. The malignant and inflammatory groups showed raised uTF levels over controls (p < 0.001). mTF levels differ similarly. For mTF and uTF assays, there was no significant difference between the malignant and inflammatory groups. The relative operating characteristic (ROC) curve plots sensitivity against false positive rate (1-specificity) for all possible cutoff values of a diagnostic test. Assay performance is assessed as the area under the curve (AUC). The ROC curve for the uTF assay displayed both sensitivity and specificity for cancer, the AUC being 0.83. Of the three mTF levels, LPS-stimulated cells gave the optimum curve (AUC = 0.71). uTF showed a weak to moderate association with mTF levels but correlated best and was statistically significant when compared with levels in the LPS-stimulated cells. uTF represents an intrinsic, kidney-derived, physiologic concentration rather than that of preactivated or postactivated monocytes. In conclusion, both uTF and LPS-stimulated mTF levels showed sensitivity and specificity in detecting cancer and inflammatory diseases. However, the two forms of TF appear to be independently derived.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Area Under Curve; Biomarkers, Tumor; Breast Neoplasms; Child; Child, Preschool; Cholelithiasis; Colorectal Neoplasms; Diagnosis, Differential; False Positive Reactions; Female; Hernia, Inguinal; Humans; Inflammation; Kidney Calculi; Lipopolysaccharides; Male; Middle Aged; Monocytes; Neoplasms; Prostatic Neoplasms; ROC Curve; Sensitivity and Specificity; Thromboplastin; Urinary Bladder Neoplasms

Recent investigations have suggested that levels of urinary tissue factor (UTF) may be elevated in some forms of cancer. We have determined UTF levels in healthy controls, patients presenting for surgery with benign prostatic hypertrophy (BPH) and untreated prostate cancer. Patients undergoing check cystoscopy, who were free of recurrent bladder cancer, and a cohort of men with bone scan positive prostate cancer recently treated by androgen ablation were also studied. UTF levels were higher in patients with prostate cancer when compared with controls, those undergoing check cystoscopy and patients with BPH. In patients with prostate cancer, bone scan positive patients had higher levels than bone scan negative subjects. The androgen ablated group had UTF levels similar to those of the control groups and significantly lower than the bone scan positive group. A weak correlation was found between UTF and serum prostate specific antigen (PSA) levels when patients with BPH and untreated cancer were analysed, but no correlation was demonstrable between PSA and UTF when cancer patients alone were evaluated. It was concluded that UTF levels are elevated in untreated prostate cancer and reflect bone scan status. In patients with bone scan positive disease UTF also reflects disease activity and may therefore be a useful disease marker in prostate cancer.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Bone and Bones; Humans; Male; Middle Aged; Prognosis; Prostate; Prostatic Hyperplasia; Prostatic Neoplasms; Radionuclide Imaging; Thromboplastin

To explore mechanisms of coagulation activation in adenocarcinoma of the prostate, the occurrence and distribution of components of coagulation and fibrinolysis pathways in situ were studied by means of immunohistochemical techniques applied to frozen sections of fresh malignant and benign hyperplastic prostatic tissue obtained at transurethral resection. Fibrinogen was distributed throughout the perivascular and tumor connective tissue in both malignant and benign disease but was not present in adjacent areas of normal prostate. Antibodies specific for fibrin and D-dimer crosslink sites stained vascular endothelium focally in both malignant and benign tissues. Both neoplastic cells and benign hyperplastic glandular epithelial cells stained weakly and in a patchy distribution for tissue factor and focally for low-molecular-weight urokinase-type plasminogen activator. Focal staining of vascular endothelium was also observed for tissue plasminogen activator and plasmin-antiplasmin complex neoantigen. By contrast, no tissue staining was observed for factor VII, factor X, factor XIII "a" subunit, high-molecular-weight urokinase-type plasminogen activator, plasminogen activator inhibitors 1 to 3, protein C, and protein S. Thus, the similarity in findings between benign hyperplastic and neoplastic prostate tissue, the lack of either an intact tumor cell-associated coagulation pathway or fibrin formation, and the presence of fibrin on vascular endothelium are consistent with the concept that coagulation activation in prostatic cancer may not be due to a direct effect of the tumor cells on the clotting mechanism. Rather, such activation may be induced by a soluble tumor product that activates procoagulant activity on certain host (for example, vascular endothelial) cells. These findings, together with the lack of effect of warfarin anticoagulation on the clinical course of patients with prostatic cancer, contrast with findings in certain other tumor types and suggest that coagulation activation may not contribute to progression of adenocarcinoma of the prostate.

Topics: Antibodies, Monoclonal; Endothelium, Vascular; Fibrin; Fibrinogen; Humans; Male; Molecular Weight; Plasminogen Activators; Prostatic Hyperplasia; Prostatic Neoplasms; Survival Rate; Thromboplastin; Warfarin

Topics: Androgen Antagonists; Disseminated Intravascular Coagulation; Humans; Male; Prostatic Neoplasms; Thromboplastin

Topics: Aged; Disseminated Intravascular Coagulation; Fibrin Fibrinogen Degradation Products; Fibrinolysis; Humans; Male; Prostatic Neoplasms; Shock, Cardiogenic; Thrombocytopenia; Thromboplastin

Topics: Aged; Blood Coagulation Tests; Blood Platelet Disorders; Chronic Disease; Disseminated Intravascular Coagulation; Fibrin; Gastrointestinal Hemorrhage; Heparin; Humans; Male; Middle Aged; Neoplasm Metastasis; Prostatic Neoplasms; Prothrombin Time; Thromboplastin

Topics: Aminocaproates; Aprotinin; Blood Platelet Disorders; Blood Platelets; Fibrinolysis; Hemorrhage; Humans; In Vitro Techniques; Male; Postoperative Complications; Prostatectomy; Prostatic Hyperplasia; Prostatic Neoplasms; Prothrombin Time; Thromboplastin