thromboplastin and Adenocarcinoma

Pulmonary tumor thrombotic microangiopathy (PTTM) is a rare clinicopathological entity causing severe pulmonary hypertension (PH). Its histological features include widespread tumor emboli of the small arteries and arterioles of the lung, associated with thrombus formation and fibrocellular and fibromuscular intimal proliferation. Although PTTM has drawn increased attention as a fatal complication of gastric carcinoma (GC), comprehensive studies are still lacking. In order to clarify clinical and pathological features of GC-induced PTTM, recent autopsy cases were analyzed with a review of the literature. Of 36 autopsy cases with GC, 6 (16.7%) were affected by PTTM. Four were male and 2 female, with a mean age of 72.7 years. Three patients presented with PTTM-related clinical manifestations and died of PTTM. They showed clear morphological evidence of PH. The other 3 patients had PTTM as an incidental finding irrespective of clinical manifestations or PH. No patient was diagnosed antemortem as PTTM. All PTTM cases were associated with advanced GC, with a histology of adenocarcinoma of poorly differentiated type (n=4) or signet-ring cell type (n=2). Expression of tissue factor and vascular endothelial growth factor was confirmed immunohistochemically in tumor cells in all cases. The results were all in line with previous studies. In addition, the current study revealed vascular lesions characteristic of PTTM morphology to be present exclusively in the lung. In conclusion, our study shows a 16.7% incidence of PTTM in GC patients, with half of them developing PH and dying of PTTM, confirming a clinical significance as a non-negligible lethal complication of GC. In addition to many known clinicopathological characteristics of PTTM, the current study pointed to some PTTM issues requiring clarification, including the pathogenesis of the exclusive pulmonary distribution of vascular lesions of PTTM. Since details remain to be elucidated, interdisciplinary research is a high priority with a close collaboration between pathologists and clinicians in order to overcome this lethal condition.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Autopsy; Carcinoma, Signet Ring Cell; Cell Differentiation; Female; Humans; Hypertension, Pulmonary; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Neoplastic Cells, Circulating; Stomach Neoplasms; Thromboplastin; Thrombotic Microangiopathies; Vascular Endothelial Growth Factor A

Cancer cells may promote fibrin formation in the tumor microenvironment through availability of procoagulant activities which are mainly of two types: tissue thromboplastin-like or direct activator of coagulation factor X. The pharmacological modulation of these activities could be potentially important in the control of metastasis growth. However, very limited information is available so far on this issue; it has recently been shown that the direct activator of coagulation factor X is a vitamin K-dependent activity which is depressed by warfarin treatment, not by anticoagulation with heparin or defibrinating enzymes. Whether the inhibition of this peculiar cancer procoagulant is involved in the antimetastatic activity of warfarin is a stimulating hypothesis which needs to be further substantiated.

Topics: Adenocarcinoma; Animals; Antineoplastic Agents; Blood Coagulation Factors; Cell Membrane; Cysteine Endopeptidases; Endopeptidases; Humans; Leukemia, Myeloid, Acute; Mice; Mucins; Neoplasm Metastasis; Neoplasm Proteins; Thromboplastin; Warfarin

Topics: Adenocarcinoma; Anemia, Hemolytic; Animals; Blood Coagulation; Disseminated Intravascular Coagulation; Erythrocyte Aging; Erythrocytes, Abnormal; Fibrinogen; Hemolytic-Uremic Syndrome; Heparin; Humans; Hypertension, Malignant; Iodine Radioisotopes; Kidney Transplantation; Liver Transplantation; Purpura, Thrombotic Thrombocytopenic; Rabbits; Thromboplastin; Transplantation, Homologous; Venoms

Limited biomarkers are used for predicting risk of venous thromboembolism (VTE) associated with cancer. Circulating microparticles (MPs), especially tissue factor- positive microparticles (TF + MPs), play an important role in the development of cancer-associated VTE. This study investigated the predictive value of plasma MPs and TF + MPs for VTE in lung cancer.. A case-control study was performed using the Beijing Chao-Yang Hospital Lung Cancer Registry. Cases had VTE occurring 3 months before or after a diagnosis of lung cancer. Controls were patients with lung cancer without VTE matched for age, histology and stage. The proportion of MPs and TF + MPs was evaluated by light-scatter-based flow cytometry.. Between January 2012 and December 2015, 30 cases with VTE and 60 controls without VTE were included. The proportion of MPs and TF + MPs was significantly more elevated in patients with VTE than in those without VTE (P < 0.05 for both). By multivariate logistic regression analysis, MPs (OR 1.153; 95% CI 1.068-1.245; P < 0.001) and adenocarcinoma (OR 3.223; 95% CI 1.062-9.782; P = 0.039) were significantly associated with VTE. The sensitivity of the proportion of MPs in diagnosing VTE was 93.3%, and the specificity was 70.0%. The sensitivity of the proportion of TF + MPs in diagnosing VTE was 66.7%, and the specificity was 88.3%. The area under the receiver operating characteristic curves for the diagnostic of the proportion of MPs and TF + MPs values were 0.836 (95% CI 0.750-0.922, P < 0.001) and 0.828 (95% CI 0.736-0.920, P < 0.001) respectively.. The elevated proportion of MPs and TF + MPs might help predict VTE associated with lung cancer.

Topics: Adenocarcinoma; Adult; Aged; Biomarkers; Case-Control Studies; Cell-Derived Microparticles; China; Female; Flow Cytometry; Humans; Lung Neoplasms; Male; Middle Aged; Plasma; Predictive Value of Tests; Retrospective Studies; Risk Assessment; Sensitivity and Specificity; Thromboplastin; Venous Thromboembolism

Topics: Adenocarcinoma; Blood Coagulation Disorders; Humans; Pancreatic Neoplasms; Poland; Thromboplastin

Topics: Adenocarcinoma; Animals; Carcinoma, Pancreatic Ductal; Immune Evasion; Mice; Mice, Inbred C57BL; Mice, Inbred NOD; Mice, SCID; Pancreatic Neoplasms; Receptor, PAR-1; Signal Transduction; Thrombin; Thromboplastin; Tumor Cells, Cultured; Tumor Microenvironment

The activation of protease-activated receptor (PAR)-2 by factor Xa (fXa) promotes the release of tissue factor-positive microvesicles (TF

Topics: Adenocarcinoma; Breast Neoplasms; Cell Line, Tumor; Cell Proliferation; Cell-Derived Microparticles; Factor VIIa; Factor Xa Inhibitors; Female; Humans; Pancreatic Neoplasms; Pyrazoles; Pyridones; Receptor, PAR-2; Rivaroxaban; Signal Transduction; Thromboplastin

Cancer patients are at high risk of developing deep venous thrombosis (DVT) and venous thromboembolism, a leading cause of mortality in this population. However, it is largely unclear how malignant tumors drive the prothrombotic cascade culminating in DVT.. Here, we addressed the pathophysiology of malignant DVT compared with nonmalignant DVT and focused on the role of tumor microvesicles as potential targets to prevent cancer-associated DVT. We show that microvesicles released by pancreatic adenocarcinoma cells (pancreatic tumor-derived microvesicles [pcMV]) boost thrombus formation in a model of flow restriction of the mouse vena cava. This depends on the synergistic activation of coagulation by pcMV and host tissue factor. Unlike nonmalignant DVT, which is initiated and propagated by innate immune cells, thrombosis triggered by pcMV was largely independent of myeloid leukocytes or platelets. Instead, we identified externalization of the phospholipid phosphatidylethanolamine as a major mechanism controlling the prothrombotic activity of pcMV. Disrupting phosphatidylethanolamine-dependent activation of factor X suppressed pcMV-induced DVT without causing changes in hemostasis.. Together, we show here that the pathophysiology of pcMV-associated experimental DVT differs markedly from innate immune cell-promoted nonmalignant DVT and is therefore amenable to distinct antithrombotic strategies. Targeting phosphatidylethanolamine on tumor microvesicles could be a new strategy for prevention of cancer-associated DVT without causing bleeding complications.

Topics: Adenocarcinoma; Animals; Bacteriocins; Blood Coagulation; Cell Line, Tumor; Cell-Derived Microparticles; Disease Models, Animal; Drug Design; Factor Xa; Fibrinolytic Agents; Humans; Mice; Mice, Inbred C57BL; Mice, Transgenic; Molecular Targeted Therapy; Pancreatic Neoplasms; Peptides; Phosphatidylethanolamines; Signal Transduction; Thromboplastin; Vena Cava, Inferior; Venous Thrombosis

The hypercoagulable state associated with pancreatic adenocarcinoma (PDA) results in increased risk of venous thromboembolism, leading to substantial morbidity and mortality. Recently, neutrophil extracellular traps (NETs), whereby activated neutrophils release their intracellular contents containing DNA, histones, tissue factor, high mobility group box 1 (HMGB1) and other components have been implicated in PDA and in cancer-associated thrombosis.. Utilizing an orthotopic murine PDA model in C57/Bl6 mice and patient correlative samples, we studied the role of NETs in PDA hypercoagulability and targeted this pathway through treatment with the NET inhibitor chloroquine. PAD4 and RAGE knockout mice, deficient in NET formation, were used to study the role of NETs in platelet aggregation, release of tissue factor and hypercoagulability. Platelet aggregation was assessed using collagen-activated impedance aggregometry. Levels of circulating tissue factor, the initiator of extrinsic coagulation, were measured using ELISA. Thromboelastograms (TEGs) were performed to assess hypercoagulability and changes associated with treatment. Correlative data and samples from a randomized clinical trial of preoperative gemcitabine/nab-paclitaxel with and without hydroxychloroquine were studied and the impact of treatment on venous thromboembolism (VTE) rate was evaluated.. The addition of NETs to whole blood stimulated platelet activation and aggregation. DNA and the receptor for advanced glycation end products (RAGE) were necessary for induction of NET associated platelet aggregation. PAD4 knockout tumor-burdened mice, unable to form NETs, had decreased aggregation and decreased circulating tissue factor. The NET inhibitor chloroquine reduces platelet aggregation, reduces circulating tissue factor and decreases hypercoagulability on TEG. Review of correlative data from patients treated on a randomized protocol of preoperative chemotherapy with and without hydroxychloroquine demonstrated a reduction in peri-operative VTE rate from 30 to 9.1% with hydroxychloroquine that neared statistical significance (p = 0.053) despite the trial not being designed to study VTE.. NETs promote hypercoagulability in murine PDA through stimulation of platelets and release of tissue factor. Chloroquine inhibits NETs and diminishes hypercoagulability. These findings support clinical study of chloroquine to lower rates of venous thromboembolism in patients with cancer.. This study reports correlative data from two clinical trials that registered with clinicaltrials.gov, NCT01128296 (May 21, 2010) and NCT01978184 (November 7, 2013).

Topics: Adenocarcinoma; Animals; Chloroquine; DNA; Extracellular Traps; Female; Humans; Hydrolases; Hydroxychloroquine; Mice; Mice, Inbred C57BL; Pancreatic Neoplasms; Platelet Aggregation; Protein-Arginine Deiminase Type 4; Receptor for Advanced Glycation End Products; Thrombelastography; Thrombophilia; Thromboplastin; Venous Thromboembolism

to study the expression of the tissue factor (TF) and its correlation with prognosis and survival in patients with gastric carcinoma.. we measured the immunohistochemical expression of TF in 50 specimens of gastric adenocarcinomas from patients submitted to curative surgery. We then compared the intensity of its expression with clinical and pathological data, TNM staging, prognostic factors and survival.. all tumors displayed TF expression; the intensity of TF expression was not associated with TNM stage, clinical or pathological variables or general survival.. TF has a high expression in gastric carcinoma, but that it is not useful as a prognostic marker.. estudar a expressão do fator tecidual (FT) e sua correlação com o prognostico e sobrevida em pacientes com carcinoma gástrico.. verificamos a expressão imuno-histoquímica do FT em 50 espécimes de adenocarcinomas gástricos de pacientes submetidos a tratamento cirúrgico com intenção curativa. A intensidade da sua expressão foi comparada com dados clínicos e patológicos, estadiamento TNM, fatores prognósticos e sobrevida.. houve expressão do FT em todos os tumores; a intensidade de expressão do FT não foi associada com estágio TNM, variáveis clínicas ou patológicas ou sobrevida geral.. este estudo mostra que o FT tem uma expressão elevada em carcinoma gástrico, mas que este não é útil como marcador de prognóstico.

Topics: Adenocarcinoma; Aged; Brazil; Female; Humans; Immunohistochemistry; Male; Middle Aged; Neoplasm Staging; Prognosis; Stomach Neoplasms; Thromboplastin

Chimeric antigen receptor (CAR)-modified T cell (CAR T) is a promising therapeutic option for patients with cancer. Such an approach requires the identification of tumor-specific antigen targets that are expressed in solid tumors. We developed a new third-generation CAR directed against tissue factor (TF), a surface molecule overexpressed in some types of lung cancer, melanoma and other cancers. First, we demonstrated by immunohistochemistry that TF was overexpressed in squamous cell carcinoma and adenocarcinoma of non-small cell lung cancer (NSCLC) and melanoma using a human tissue microarray. In the presence of TF-positive cancer cells, the CAR-modified T cells (TF-CAR T) were highly activated and showed specific cytotoxicity to TF-positive cancer cells in vitro. In established s.c. xenograft and lung metastasis models, TF-CAR T cells could significantly suppress the growth of s.c. xenograft and metastasis of TF-positive cancer cells. Additionally, the safety evaluation of TF-CAR T cells in vivo showed that the treatment did not cause obvious toxicity in mice. Taken together, these findings indicate that TF-CAR T cells might be a novel potential therapeutic agent for the treatment of patients with TF-positive cancers.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Animals; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Cell Movement; Cell Proliferation; Cytokines; Cytotoxicity, Immunologic; Female; Humans; Immunotherapy, Adoptive; Lung Neoplasms; MCF-7 Cells; Melanoma, Experimental; Mice, Inbred NOD; Mice, SCID; Neoplasm Invasiveness; Receptors, Antigen, T-Cell; Skin Neoplasms; T-Lymphocytes; Thromboplastin; Time Factors; Transfection; Tumor Burden; Xenograft Model Antitumor Assays

ESSENTIALS: Cancer patients have a high rate of venous thrombosis (VT) but the underlying mechanisms are unknown. Tumor-derived, tissue factor-positive microvesicles in platelet activation in vitro and in vivo were studied. Tumor-derived, tissue factor-positive microvesicles enhanced VT in mice. Platelets may contribute to VT in some cancer patients, and this could be prevented with antiplatelet drugs.. Cancer patients have an approximately 4-fold increased risk of venous thromboembolism (VTE) compared with the general population, and cancer patients with VTE have reduced survival. Tumor cells constitutively release small membrane vesicles called microvesicles (MVs) that may contribute to thrombosis in cancer patients. Clinical studies have shown that levels of circulating tumor-derived, tissue factor-positive (TF(+) ) MVs in pancreatic cancer patients are associated with VTE. Objectives We tested the hypothesis that TF(+) tumor-derived MVs (TMVs) activate platelets in vitro and in mice.. We selected two human pancreatic adenocarcinoma cell lines expressing high (BxPc-3) and low (L3.6pl) levels of TF as models to study the effect of TF(+) TMVs on platelets and thrombosis.. We found that both types of TF(+) TMVs activated human platelets and induced aggregation in vitro in a TF and thrombin-dependent manner. Further, injection of BxPc-3 TF(+) TMVs triggered platelet activation in vivo and enhanced thrombosis in two mouse models of venous thrombosis in a TF-dependent manner. Importantly, BxPc-3 TF(+) TMV-enhanced thrombosis was reduced in Par4-deficient mice and in wild-type mice treated with clopidogrel, suggesting that platelet activation was required for enhanced thrombosis. These studies suggest that TF(+) TMV-induced platelet activation contributes to thrombosis in cancer patients.

Topics: Adenocarcinoma; Animals; Blood Platelets; Cell Line, Tumor; Cell-Derived Microparticles; Clopidogrel; Female; Flow Cytometry; Humans; Mice; Mice, Inbred C57BL; Neoplasms; Pancreatic Neoplasms; Platelet Activation; Platelet Aggregation; Platelet Aggregation Inhibitors; Pulmonary Embolism; Thrombin; Thromboplastin; Thrombosis; Ticlopidine

Highly elevated microparticle (MP)-associated tissue factor (TF) activity was found in patients with pancreatic cancer, one of the most prothrombotic malignancies. It remains to be elucidated whether MP-TF activity reflects the prothrombotic state in these patients. MP-TF activity levels and the TF-dependent and -independent effect of MPs on fibrin clot formation were determined in patients with metastatic pancreatic cancer (n = 27), in healthy individuals (n = 10) and in plasma samples from lipopolysaccharide (LPS)-stimulated blood (LPS-plasma), which is rich in monocyte-derived TF-bearing MPs. The median MP-TF activity was 1.06 pg/mL (range, from 0.19 to 10.34 pg/mL) in patients with pancreatic cancer, 0.61 pg/mL (range, from 0.36 to 0.79 pg/mL) in LPS-plasma, and 0.18 pg/mL (range, from 0.04 to 0.39 pg/mL) in healthy individuals. MPs derived from LPS-plasma had the strongest impact on fibrin clot formation time (median, 157.6 seconds; range, from 149.5 to 170.4 seconds). Fibrin clot formation occurred significantly later in MPs derived from patients with pancreatic cancer (median, 273.4 seconds; range, from 146.6 to 354.4 seconds; P < 0.001) and in healthy individuals (median, 299.0 seconds; range, from 261.1 to 417.9 seconds; P < 0.001). Only in MPs derived from LPS-plasma the fibrin clot formation time dependent strongly on TF (median prolongation after TF blockade: 68% in LPS-plasma, 10% in patients with pancreatic cancer, and 4% in healthy individuals). In conclusion, highly elevated MP-TF activity was found in patients with metastatic pancreatic cancer, but TF-bearing MPs had a small effect on fibrin clot formation. TF-bearing MPs might not be the main mediators of the prothrombotic state associated with pancreatic cancer. However, the small but significant increase in coagulation potential by TF-bearing MPs might contribute to the multifactorial pathogenesis of venous thromboembolism in pancreatic cancer.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Case-Control Studies; Female; Fibrin; Humans; Male; Middle Aged; Neoplasm Metastasis; Pancreatic Neoplasms; Thromboplastin; Thrombosis

Topics: Adenocarcinoma; Female; Fibrin; Humans; Male; Neoplasm Metastasis; Pancreatic Neoplasms; Thromboplastin; Thrombosis

In this study, 52 patients were studied to elucidate the relative impact of resection of localized pancreaticobiliary adenocarcinoma (PBC) on circulating factors of tumour-associated angiogenesis e.g. tissue factor bearing microparticles (TFMP) and vascular endothelial growth factor (VEGF) and their clinicopathological significance to angiogenesis markers in cancer tissue from PBC patients. Angiogenesis array analysis on serum samples revealed that surgical resection of tumour lesion in PBC patients affects the levels of a panel of angiogenesis-related molecules, including VEGF that was verified by ELISA to significantly reduce (median & IQR: 1003(369-2000) vs. 457(159-834) pg/ml; p<0.05). Correspondingly, a significant decrease in the angiogenic activity (decreased capillary tube formation; p<0.05) of serum samples after the surgery was also found. Despite a decrease in number of circulating TFMP after surgery, this did not reach statistical significance; there was a significant reduction in pro-coagulant activity (prolonged prothrombin time, p<0.001) post-operatively. In addition, the activity of total microparticles (MP activity assay, p<0.05) was decreased significantly. Immunohistochemical staining of tumour tissue revealed a strong correlation between the microvessel density (MVD) and VEGF expression. Also, higher levels of circulating TFMP or TF related activity (prothrombin time) correlated significantly with TF expression and MVD on tumour tissues from PBC patients. These findings suggest that in pancreaticobiliary adenocarcinoma TF related angiogenesis drivers are equally significant to VEGF ones, raising the clinical question of whether the effectiveness of angiogenesis targeting studies could be improved through the 'dual' targeting of these pathways in PBC.

Topics: Adenocarcinoma; Aged; Cell-Derived Microparticles; Female; Humans; Male; Middle Aged; Neovascularization, Pathologic; Pancreas; Pancreatic Neoplasms; Thromboplastin; Thrombosis; Vascular Endothelial Growth Factor A

Malignant pleural effusion (MPE) is a poor prognostic sign for patients with lung cancer. Tissue factor (TF) is a coagulation factor that participates in angiogenesis and vascular permeability and is abundant in MPE. We previously demonstrated that autocrine IL-6-activated Stat3 contributes to tumor metastasis and upregulation of VEGF, resulting in the generation of MPE in lung adenocarcinoma. In this study, we found IL-6-triggered Stat3 activation also induces TF expression. By using pharmacologic inhibitors, it was shown that JAK2 kinase, but not Src kinase, contributed to autocrine IL-6-induced TF expression. Inhibition of Stat3 activation by dominant negative Stat3 (S3D) in lung adenocarcinoma suppressed TF-induced coagulation, anchorage-independent growth in vitro, and tumor growth in vivo. Consistently, knockdown of TF expression by siRNA resulted in a reduction of anchorage-independent growth of lung adenocarcinoma cells. Inhibition of TF expression also decreased the adhesion ability of cancer cells in normal lung tissues. In the nude mouse model, both lung metastasis and MPE generation were decreased when PC14PE6/AS2-siTF cells (TF expression was silenced) were intravenously injected. PC14PE6/AS2-siTF cells also produced less malignant ascites through inhibition of vascular permeability. In summary, we showed that TF expression plays a pivotal role in the pathogenesis of MPE generation via regulating of tumor metastasis and vascular permeability in lung adenocarcinoma bearing activated Stat3.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Animals; Blotting, Western; Capillary Permeability; DNA Primers; Flow Cytometry; Fluorescent Antibody Technique; Gene Knockdown Techniques; Interleukin-6; Janus Kinase 2; Luciferases; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Metastasis; Pleural Effusion, Malignant; Reverse Transcriptase Polymerase Chain Reaction; RNA, Small Interfering; STAT3 Transcription Factor; Thromboplastin; Up-Regulation

Patients with lung adenocarcinoma undergoing surgery are in high risk for VTE and receive routine post-operative thromboprophylaxis with LWMH.. We investigated markers of hypercoagulability in patients with primary localized adenocarcinoma and the modifications induced by lobectomy and postoperative administration of enoxaparin.. Patients suffering from localised primary lung adenocarcinoma (n=15) scheduled for lobectomy were studied. The control group consisted of 15 healthy age and sex-matched individuals. Blood was collected before anaesthesia induction and after surgery, at several intervals until the 7th post-operative day. Samples were assessed for thrombin generation, phosphatidylserin expressing platelet derived microparticles expressing (Pd-MP/PS(+)), tissue factor activity (TFa), FVIIa and TFPI levels, procoagulant phospholipid dependent clotting time and anti-Xa activity.. At baseline, patients showed increased thrombin generation and Pd-MP/PS(+). After lobectomy thrombin generation significantly decreased. Administration of enoxaparin attenuated thrombin generation. In about 50% of samples collected post-operatively an increase of thrombin generation occurred despite the presence of the expected anti-Xa activity in plasma. At the 7th post-operative day, 3 out of 15 patients showed a significant increase of thrombin generation.. In patients with localized lung adenocarcinoma, hypercoagulability is characterized by high thrombin generation and increased concentration of Pd-MP/PS(+). Tumor mass resection is related with attenuation of thrombin generation, which is inhibited by postoperative thromboprophylaxis with enoxaparin. The response to enoxaparin is not predicted by the concentration of the anti-Xa activity in plasma. The assessment of thrombin generation during prophylaxis with enoxaparin allows to identify patients with high residual plasma hypercoagulability.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Aged; Anticoagulants; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Cell-Derived Microparticles; Enoxaparin; Factor Xa Inhibitors; Female; Humans; Lung; Lung Neoplasms; Male; Middle Aged; Postoperative Period; Thrombin; Thrombophilia; Thromboplastin

Patients with pancreatic cancer have an unfavourable prognosis. A central role in pancreatic cancer progression has been suggested for tissue factor (TF), the main initiator of the blood coagulation cascade. We hypothesized that elevated levels of plasma microparticle (MP)-associated TF activity might indicate the presence of poorly differentiated pancreatic cancer, disease dissemination and infiltration of peripancreatic vessels.. MP-TF activity was measured in 73 pancreatic cancer patients and 22 healthy controls. Abdominal computerized tomography (CT) scans performed at study inclusion were investigated for probability of tumoural vascular invasion. In addition, intratumoural TF expression, D-dimer and CA 19-9 levels were determined.. MP-TF activity (pg/mL) was significantly higher in patients (median: 0·37 [range: 0·00-11·91]) than in controls (median: 0·05 [range: 0·00-0·76]; P < 0·001). When pancreatic cancer patients were compared with regard to stage and grade, significantly elevated levels of MP-TF activity were only present in those with poorly differentiated metastatic nonresectable tumours (n = 11, median: 2·95 [range: 0·25-11·91]). In three patients with poorly differentiated tumours, a high probability of vascular invasion was found (MP-TF activity in these cases: 2·95, 7·00 and 10·34). MP-TF activity correlated strongly with CA 19-9 (r = 0·60) and weakly with D-dimer (r = 0·33) levels. Immunohistochemical staining for TF was positive in 14 of 15 resected tumours. MP-TF activity was associated with an increased risk of mortality (HR: 1·8 per doubling in MP-TF activity, [95% CI: 1·4-2·4, P < 0·001]).. MP-TF activity might represent a biomarker for a poorly differentiated and invasive pancreatic cancer phenotype and poor survival.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; CA-19-9 Antigen; Cell-Derived Microparticles; Female; Fibrin Fibrinogen Degradation Products; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Male; Middle Aged; Neoplasm Grading; Neoplasm Invasiveness; Neoplasm Staging; Pancreatic Neoplasms; Prognosis; Thromboplastin; Tomography, Spiral Computed; Vascular Neoplasms

The overexpression of tissue factor (TF) observed in numerous cancer cells and clinical samples of human cancers make TF an ideal target for cancer therapy. Here, we report an energized fusion protein, hlFVII-LDP-AE, which can be used for cancer therapy and is composed of a human Factor VII light chain (hlFVII) conjugated to the cytotoxic antibiotic lidamycin (LDM, LDP-AE). hlFVII-LDP-AE binds with specificity to TF expressed on tumor cells, resulting in internalization of the fusion protein and cytotoxicity induced by the LDM domain. The potential efficacy of hlFVII-LDP-AE for cancer therapy was examined in vitro by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and in vivo with a BALB/c nude mouse xenograft model of the human lung cancer line NCI-H292. hlFVII-LDP-AE caused chromatin condensation and cleavage of genomic DNA in NCI-H292 cells. In the MTT assays, the IC50 value of hlFVII- LDP-AE was 0.19 nM. In the in vivo tests, after two intravenous injections of hlFVII-LDP-AE at a dose of 0.6 mg/kg, the growth rate of the lung tumor xenograft was reduced to 15% of the control rate, and there was no excessive loss of body weight and inflammatory response in the mice. These findings suggest that hlFVII-LDP-AE is efficacious and tolerated in the mouse model of NCI-H292 human lung cancer examined and could have broad clinical applicability for treating cancer patients.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Aminoglycosides; Animals; Antineoplastic Agents; Cell Death; Cell Line, Tumor; Cell Proliferation; Chromatin Assembly and Disassembly; DNA Damage; Enediynes; Factor VII; Female; Humans; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Protein Binding; Recombinant Fusion Proteins; Thromboplastin; Xenograft Model Antitumor Assays

The renin-angiotensin system (RAS) promotes angiogenesis and growth of neoplastic cells. Angiotensin-converting enzyme (ACE) inhibitors and angiotensin II receptor AT1 blockers may protect against cancer. Tissue factor (TF), for its involvement in tumor growth, angiogenesis, and metastasis is considered a hallmark of cancer progression. In this study we evaluated whether RAS blockade modulates TF constitutive expression by the metastatic breast carcinoma MDA-MB-231 cell line.. Cell TF activity was assessed by one stage clotting time, TF and VEGF antigens and mRNA levels by ELISA and RT-PCR, respectively. AT(1) was detected by flow-cytometry and angiotensin-II levels by EIA.. Captopril reduced in a concentration-dependent way both the strong constitutive TF activity (983.2±55.2 vs. 686.7±135.1U/5×10(5) cells with 10μg/ml captopril) and antigen (32.3±5.9 vs. 13.2±6.6ng/ml) in MDA-MB-231. Similar results were observed with enalapril. AT1 was present on cell membrane and losartan, a competitive inhibitor of AT1, reduced TF expression to a degree similar as that exerted by ACE inhibitors. Moreover, captopril and losartan downregulated the constitutive mRNA TF expression by ~35%. Similar results were observed with anti-AT1 and angiotensin II antibodies. In addition, the constitutive VEGF antigen and mRNA levels were reduced in the presence of captopril or losartan, and an anti-VEGF antibody downregulated cell TF activity by ~40%.. These results could, at least in part, contribute to the discussion about the possible effects of ACE inhibitors and AT1 receptor antagonists in malignancy, and offer new clues to support their use for tumor control.

Topics: Adenocarcinoma; Angiotensin-Converting Enzyme Inhibitors; Breast Neoplasms; Captopril; Cell Line, Tumor; Down-Regulation; Enalapril; Female; Humans; MCF-7 Cells; Neovascularization, Pathologic; Renin-Angiotensin System; Thromboplastin; Vascular Endothelial Growth Factor A

Tissue factor (TF), which has a role in normal tissue haemostasis, was reported to be aberrantly expressed, associated with higher microvascular density and a poor prognosis in intestinal-type gastric adenocarcinoma in the Japanese population. This is the first study to look at the relationship of TF and the metaplasia-adenoma-carcinoma sequence (MACS) of gastric cancer in a European population.. The expression of TF was examined immunohistochemically in 191 gastric tissue samples: (13: normal; 18: intestinal metaplasia; 160: gastric adenocarcinoma) from the European population.. TF was not expressed in normal gastric mucosal cells. A strong intensity of staining was found in intestinal metaplasia cells but in 2 of 18 samples. TF expression increased with advancing stage of gastric cancer (P<0.0001, Jonckheere's test for ordered medians). Stage 3-4 gastric cancers preferentially expressed TF (34%, P=0.04). In comparison with the Japanese study, TF was not expressed at a higher level in intestinal vs diffuse-type gastric cancers and expression had 'no prognostic' significance.. TF may be involved in tumour progression along the MACS of gastric cancer in the European population and is shown to start in precancerous lesions. However, clinical features may differ due to differences in biological features in the two populations, as reflected by differences in TF expression profile.

Topics: Adenocarcinoma; Adenoma; Adult; Aged; Aged, 80 and over; Carcinoma; Disease Progression; Female; Gastric Mucosa; Humans; Immunohistochemistry; Male; Metaplasia; Middle Aged; Prognosis; Stomach; Stomach Neoplasms; Thromboplastin; White People

Pulmonary tumor thrombotic microangiopathy (PTTM) has been known as a rare and serious cancer-related pulmonary complication. However, the pathogenesis and pathophysiology of this debilitating condition still remains obscure and no effective management was recommended. The present study aims to elucidate the pathophysiology of PTTM.. Autopsy records were searched to extract cases of pulmonary tumor embolism induced by metastasis of gastric carcinoma in the Toho University Omori Medical Center from 2000 to 2006. And then, tissue sections of extracted cases were prepared for not only light microscopic observation but morphometric analysis with the use of selected PTTM cases.. Six autopsies involved PTTM and clinicopathological data of them were summarized. There was a significant negative association between pulmonary arterial diameter and stenosis rate in four cases. Although all cases showed an increase of stenosis rate to some degree, the degree of stenosis rate varied from case to case. Significant differences were found for average stenosis rate between the under 100 micrometer group or the 100 to 300 micrometer group and the 300 micrometer group in four cases. However, no significant differences were found for average stenosis rate between the under 100 micrometer group and the 100 to 300 micrometer group in all cases. Meanwhile, all cases showed positive reactivity for tissue factor (TF), five showed positive reactivity for vascular endothelial growth factor (VEGF), and three showed positive reactivity for osteopontin (OPN).. In the present study, we revealed that the degree of luminal narrowing of the pulmonary arteries varied from case to case, and our results suggested that pulmonary hypertension in PTTM occurs in selected cases which have a widespread pulmonary lesion with severe luminal narrowing in the smaller arteries. Furthermore, our immunohistochemical examination indicated that gastric carcinoma indicating PTTM shows a higher TF-positive rate than typical gastric carcinoma. However, it remains still obscuring whether gastric carcinoma indicating PTTM shows a higher VEGF or OPN-positive rate as determined by immunohistochemistry.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Biomarkers, Tumor; Fatal Outcome; Female; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Neoplastic Cells, Circulating; Osteopontin; Pulmonary Artery; Stomach Neoplasms; Thromboplastin; Thrombotic Microangiopathies; Vascular Endothelial Growth Factor A

The human coagulation trigger tissue factor (TF) is overexpressed in several types of cancer and involved in tumor growth, vascularization, and metastasis. To explore the role of TF in biological processes of lung adenocarcinoma, we used RNA interference (RNAi) technology to silence TF in a lung adenocarcinoma cell line A549 with high-level expression of TF and evaluate its antitumor effects in vitro and in vivo.. The specific small interfering RNA (siRNA) designed for targeting human TF was transfected into A549 cells. The expression of TF was detected by reverse transcription-PCR and Western blot. Cell proliferation was measured by MTT and clonogenic assays. Cell apoptosis was assessed by flow cytometry. The metastatic potential of A549 cells was determined by wound healing, the mobility and Matrigel invasion assays. Expressions of PI3K/Akt, Erk1/2, VEGF and MMP-2/-9 in transfected cells were detected by Western blot. In vivo, the effect of TF-siRNA on the growth of A549 lung adenocarcinoma xenografts in nude mice was investigated.. TF -siRNA significantly reduced the expression of TF in the mRNA and protein levels. The down-regulation of TF in A549 cells resulted in the suppression of cell proliferation, invasion and metastasis and induced cell apoptosis in dose-dependent manner. Erk MAPK, PI3K/Akt pathways as well as VEGF and MMP-2/-9 expressions were inhibited in TF-siRNA transfected cells. Moreover, intratumoral injection of siRNA targeting TF suppressed the tumor growth of A549 cells in vivo model of lung adenocarcinoma.. Down-regulation of TF using siRNA could provide a potential approach for gene therapy against lung adenocarcinoma, and the antitumor effects may be associated with inhibition of Erk MAPK, PI3K/Akt pathways.

Topics: Adenocarcinoma; Adenocarcinoma of Lung; Animals; Apoptosis; Cell Line, Tumor; Cell Movement; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; RNA, Small Interfering; Signal Transduction; Thromboplastin; Tumor Burden; Xenograft Model Antitumor Assays

Cervical cancer continues to be an important worldwide health problem for women. Up to 35% of patients who are diagnosed with and appropriately treated for cervical cancer will recur and treatment results are poor for recurrent disease. Given these sobering statistics, development of novel therapies for cervical cancer remains a high priority. We evaluated the expression of Tissue Factor (TF) in cervical cancer and the potential of hI-con1, an antibody-like-molecule targeted against TF, as a novel form of immunotherapy against multiple primary cervical carcinoma cell lines with squamous- and adenocarcinoma histology.. Because TF is a transmembrane receptor for coagulation factor VII/VIIa (fVII), in this study we evaluated the in vitro expression of TF in cervical carcinoma cell lines by immunohistochemistry (IHC), real time-PCR (qRT-PCR) and flow cytometry. Sensitivity to hI-con1-dependent cell-mediated-cytotoxicity (IDCC) was evaluated in 5-hrs-51chromium-release-assays against cervical cancer cell lines in vitro.. Cytoplasmic and/or membrane TF expression was observed in 8 out of 8 (100%) of the tumor tissues tested by IHC and in 100% (11 out of 11) of the cervical carcinoma cell lines tested by real-time-PCR and flow cytometry but not in normal cervical keratinocytes (p=0.0023 qRT-PCR; p=0.0042 flow cytometry). All primary cervical cancer cell lines tested overexpressing TF, regardless of their histology, were highly sensitive to IDCC (mean killing±SD, 56.2%±15.9%, range, 32.4%-76.9%, p<0.001), while negligible cytotoxicity was seen in the absence of hI-con1 or in the presence of rituximab-control-antibody. Low doses of interleukin-2 further increased the cytotoxic effect induced by hI-con1 (p=0.025) while human serum did not significantly decrease IDCC against cervical cancer cell lines (p=0.597).. TF is highly expressed in squamous and adenocarcinoma of the uterine cervix. hI-con1 induces strong cytotoxicity against primary cervical cancer cell lines overexpressing TF and may represent a novel therapeutic agent for the treatment of cervical cancer refractory to standard treatment modalities.

Topics: Adenocarcinoma; Carcinoma, Squamous Cell; Cell Line, Tumor; Complement System Proteins; Cytotoxicity Tests, Immunologic; Drug Screening Assays, Antitumor; Female; Human papillomavirus 16; Human papillomavirus 18; Humans; Immunoconjugates; Immunoglobulin G; Immunotherapy; Interleukin-2; Keratinocytes; Molecular Targeted Therapy; Neoplasm Proteins; Neovascularization, Pathologic; Papillomavirus Infections; RNA, Messenger; RNA, Neoplasm; Thromboplastin; Uterine Cervical Neoplasms

Advanced pancreatic cancer is associated with a high risk of patients developing venous thromboembolism. This increased risk is thought to be tumour-driven and associated with tissue factor (TF) and microparticles. The aim of this study was to investigate the role of TF and phospholipid expression in the procoagulant properties of pancreatic cell lines and microparticles. Pancreatic cancer cell lines (MIA-PaCa-2, ASPC-1 and CFPAC-1) were assessed for expression of TF and microparticle release. Procoagulant potential was determined by a prothrombin time assay. Cell surface expression of TF was highest in CFPAC-1, with low expression on ASPC-1 and little/no expression on MIA-PaCa-2. Clotting time (CT) was cell number and TF-dependent (P < 0.001). Blocking of TF resulted in slower CT for CFPAC-1 and ASPC1 and prevented clotting in MIA-PaCa-2. Microparticles were shown to be procoagulant and the majority of procoagulant potential could be removed by passing cell-free media through a 0.1 μm filter. A dose-dependent CT was observed in both ASPC-1 and CFPAC-1 cell-free media. Furthermore, addition of duramycin prevented microparticle-supported coagulation. The data presented suggest a key role for cell and microparticle surface-expressed TF and phospholipids in coagulation and highlight duramycin-mediated disruption of clotting.

Topics: Adenocarcinoma; Adult; Aged; Bacteriocins; Blood Coagulation; Blood Coagulation Tests; Carcinoma; Cell Line, Tumor; Cell-Derived Microparticles; Culture Media, Conditioned; Female; Humans; Male; Middle Aged; Organ Specificity; Pancreas; Pancreatic Neoplasms; Peptides; Phosphatidylethanolamines; Phosphatidylserines; Prothrombin Time; Thrombin; Thromboplastin; Venous Thromboembolism

Tissue factor (TF)-mediated protease-activated receptor (PAR)-2 signaling is associated with a promigratory, invasive and proangiogenic phenotype in experimental models of breast cancer and has been mechanistically coupled to phosphorylation of the TF cytoplasmic domain (pTF). However, the clinical relevance of these findings is unknown. Here, we provide the first in vivo evidence of TF phosphorylation in experimental as well as clinical breast cancer tumors. pTF was demonstrated in MDA-MB-231 xenografts and in tumors from the MMTV-PyMT transgene model of spontaneous murine breast adenocarcinoma. Tumors from PAR-2-deficient transgenic mice were negative for pTF, thus linking pTF to PAR-2 signaling. The clinical correlation between TF, pTF, PAR-1, PAR-2 and vascular endothelial growth factor (VEGF)-A was determined by immunohistochemistry on tumors from a cohort of 172 consecutive primary breast cancer patients, with a median follow-up time of 50 months. In 160 evaluable patient tumors, pTF was associated with TF (p = 0.01) and cancer cell expression of PAR-1 (p = 0.001), PAR-2 (p = 0.014) and VEGF-A (p = 0.003) using chi(2) test. PAR-2 and VEGF-A were coexpressed (p = 0.013) and associated with a more aggressive phenotype. Interestingly, all patients experiencing recurrences had tumors expressing pTF and PAR-2, and pTF alone as well as coexpression of pTF and PAR-2 were significantly correlated with shorter recurrence-free survival (log rank test, p = 0.04 and p = 0.02, respectively). This study provides the first evidence to link PAR-2 expression and TF phosphorylation to clinical data in human breast cancer. In conjunction with experimental tumor models, these data support an important role of TF-PAR-2 signaling in breast cancer recurrence.

Topics: Adenocarcinoma; Adult; Animals; Breast Neoplasms; Cell Line, Tumor; Female; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Lymphatic Metastasis; Mammary Neoplasms, Animal; Mammary Neoplasms, Experimental; Mice; Mice, SCID; Middle Aged; Neoplasm Transplantation; Phosphorylation; Predictive Value of Tests; Prognosis; Prospective Studies; Receptor, PAR-2; Signal Transduction; Sweden; Thromboplastin; Transplantation, Heterologous; Vascular Endothelial Growth Factor A

A correlation between cancer and hypercoagulability has been described for more than a century. Patients with cancer are at increased risk for thrombotic complications and the clotting initiator protein, tissue factor (TF), is possibly involved in this process. Moreover, TF may promote angiogenesis and tumor growth. In addition to TF, thrombin seems to play a relevant role in tumor biology, mainly through activation of protease-activated receptor-1 (PAR-1). In the present study, we prospectively studied 39 lung adenocarcinoma patients in relation to the tumor expression levels of TF and PAR-1 and their correlation with thrombosis outcome and survival. Immunohistochemical analysis showed TF positivity in 22 patients (56%), most of them in advanced stages (III and IV). Expression of PAR-1 was found in 15 patients (39%), most of them also in advanced stages (III and IV). Remarkably, no correlation was observed between the expression of TF or PAR-1 and risk for thrombosis development. On the other hand, patients who were positive for TF or PAR-1 tended to have decreased long-term survival. We conclude that immunolocalization of either TF or PAR-1 in lung adenocarcinoma may predict a poor prognosis although lacking correlation with thrombosis outcome.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Female; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Neoplasm Seeding; Prognosis; Prospective Studies; Receptor, PAR-1; Thromboplastin; Thrombosis

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

Non-small cell lung cancer (NSCLC) comprises of 75% of all lung cancers. Human full length tissue factor (flHTF), the physiological initiator of blood coagulation, is aberrantly expressed in certain solid tumors. FlHTF and its soluble isoform, alternatively spliced human tissue factor (asHTF), have been shown to contribute to thrombogenicity of the blood of healthy individuals. The aim of this study was to quantify flHTF and asHTF on mRNA and protein levels (using immunohistochemistry, immunoblotting, and ELISA) on a panel of human NSCLC tissue and plasma specimens. The tissue factor (TF) expression of 21 pulmonary adenomatous (AC) and 12 normal healthy tissues was assessed by real-time qRT-PCR. The TF protein concentration was quantified by ELISA in a subset of 11 AC and 9 normal tissue specimens as well as in the plasma of 13 lung cancer patients and 15 healthy controls. We found a significant increase in the ratio of flHTF/HGAPDH mRNA in AC (0.24+/-0.06 vs. 0.07+/-0.01; p=0.02 vs. controls) and in asHTF/HGAPDH mRNA (0.027+/-0.01 vs. 0.004+/-0.001; p=0.03 AC vs. controls). AsHTF mRNA expression was significantly lower in patients with stage IA disease compared to patients with higher grade stages, pointing to TF as being a marker of malignancy and metastases. TF protein of lung tumors was significantly increased in AC (p=0.004 vs. controls). TF in plasma was up-regulated in lung cancer patients (334.9+/-95.4 vs. 124.1+/-14.8 pg/ml; p=0.02 vs. controls). Immunohistochemical and immunoblotting data are in line with the increased TF expression, showing elevated blood thrombogenicity of NSCLC patients. The up-regulation of flHTF and, especially, asHTF in AC suggests not only a raised risk of thrombosis, but also of tumor progression, thereby, indicating a poor prognosis in these patients.

Topics: Adenocarcinoma; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Humans; Immunohistochemistry; Lung Neoplasms; Neoplasm Metastasis; RNA, Messenger; Thromboplastin; Thrombosis

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

Tissue factor (TF), which normally safeguards vascular integrity by inducing hemostasis upon injury, has received widespread attention in the pathogenesis of cancer progression and metastasis. Aberrantly expressed TF in cancer cells has been reported to be associated with advanced stages of malignancy in various cancers.. The expression of TF and microvessel density (MVD) were immunohistochemically evaluated in 207 gastric cancers, and their relationship with clinicopathological features was examined.. TF was preferentially expressed (41.8%) in intestinal-type cancer at a significantly higher rate than that in diffuse-type cancer (12.1%, P<0.0001). The expression of TF was associated with advanced stage of disease and showed a positive correlation with a higher rate of lymphatic and venous invasion and lymphatic metastasis in intestinal-type, but not in diffuse-type carcinoma. Moreover, TF expression was associated with high MVD in the tumor and a worse outcome only in intestinal-type carcinoma.. TF may be critically involved in tumor progression in intestinal-type, but not in diffuse-type, gastric carcinoma. The difference in clinical features between these two histological types might be partially dependent on TF expression profile.

Topics: Adenocarcinoma; Aged; Carcinoma, Signet Ring Cell; Female; Gastrectomy; Humans; Intestinal Neoplasms; Lymph Node Excision; Lymph Nodes; Lymphatic Metastasis; Male; Microcirculation; Middle Aged; Neoplasm Invasiveness; Prognosis; Stomach Neoplasms; Thromboplastin

The aim of this study was to examine the relationship between tissue factor (TF), vascular endothelial growth factor (VEGF) and the onset of angiogenesis in the adenoma-carcinoma sequence (ACS), the stepwise process encompassing colorectal cancer (CRC) disease progression.. 210 surgical specimens comprising the ACS were immunohistochemically stained for endothelial cells (CD31), VEGF and TF. Angiogenesis quantified using Chalkley grid analysis (microvascular density; MVD), and VEGF/TF expression were semiquantitatively graded and correlated with standard prognostic indicators including 5 year follow-up. VEGF and TF were measured by ELISA in tumour specimens and normal mucosa from an additional 90 CRC patients.. There was a significant increase in MVD across the ACS (p < 0.0005) with significant correlations with Dukes' stage (p = 0.01) and lymph node involvement (p = 0.02). The greatest increase in MVD was related to the onset of dysplasia, with an associated significant increase in VEGF expression (p < 0.0005). There was a significant relationship between VEGF and TF expression in the initial phase of the ACS (k = 0.44, p < 0.005), although no correlation between VEGF or TF, and MVD, tumour size, Dukes' classification, lymph node involvement or survival was found.. These findings are the first to suggest that the angiogenic switch occurs at the onset of dysplasia in the ACS, and provide further evidence of the close association between VEGF and TF in the early stages of CRC development.

Topics: Adenocarcinoma; Adenoma; Adult; Aged; Aged, 80 and over; Colorectal Neoplasms; Disease Progression; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Male; Middle Aged; Neoplasm Proteins; Neovascularization, Pathologic; Survival Analysis; Thromboplastin; Vascular Endothelial Growth Factor A

Tissue factor (TF) is a transmembrane glycoprotein that serves as the prime initiator of blood coagulation and plays a critical role in thrombosis and hemostasis. In addition, a variety of tumor cells overexpress cell-surface TF, which appears to be important for tumor angiogenesis and metastasis. To elucidate the mechanism involved in the upregulation of TF in human tumor cells, a comprehensive analysis of TF mRNA from various normal and tumor cells was performed. The results of these studies indicate that, in addition to possessing a normal full-length TF transcript and minor levels of an alternatively spliced transcript known as alternatively-spliced tissue factor (asTF), human tumor cells express additional full-length TF transcripts that are also generated by alternative splicing. Reverse transcriptase-polymerase chain reaction (RT-PCR) and 5'-rapid amplification of cDNA ends- (5'-RACE) based analyses of cytoplasmic RNA from normal and tumor cells revealed that there is alternative splicing of the first intron between exon I and exon II resulting in 2 additional TF transcripts. One of the transcripts has an extended exon I with inclusion of most of the first TF intron (955 bp), while the second transcript is formed by the insertion of a 495 bp sequence, referred to as exon IA, derived from an internal sequence of the first intron. The full length TF transcript with alternatively spliced novel exon IA, referred to as alternative exon 1A-tissue factor (TF-A), represented approximately 1% of the total TF transcripts in normal cells, but constituted 7-10% of the total TF transcript in tumor cells. Quantitative real-time RT-PCR analysis indicated that cultured human tumor cells contain 10-25-fold more copy numbers of TF-A in comparison to normal, untransformed cells. We propose that high-level expression of the novel TF-A transcript, preferentially in tumor cells, may have utility in the diagnosis and staging of a variety of solid tumors.

Topics: Adenocarcinoma; Alternative Splicing; Base Sequence; Carcinoma, Hepatocellular; Carcinoma, Transitional Cell; Cytoplasm; Exons; Humans; Introns; Leukemia, Promyelocytic, Acute; Liver Neoplasms; Molecular Sequence Data; Neoplasm Staging; Neoplasms; Pancreatic Neoplasms; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Thromboplastin; Tumor Cells, Cultured; Up-Regulation

To study expression of tissue factor (TF) in pancreatic cancer and its role in the development of thromboembolism.. TF expression was studied in eight human pancreatic carcinoma cell lines by Northern blot and indirect immunofluorescence. Expression of alternatively spliced TF (asTF) was assessed by RT-PCR. In addition, TF expression was determined by immunofluorescence in pancreatic tissues of 19 patients with pancreatic adenocarcinoma (PCa), 9 patients with chronic pancreatitis (CP) and 20 normal controls. Plasma samples (30 PCa-patients, 13 CP-patients and 20 controls) were investigated for soluble TF levels and coagulation activation markers [thrombin-antithrombin III complex (TAT), prothrombin fragment 1 + 2 (F1 + 2)].. All pancreatic carcinoma cell lines expressed TF (8/8) and most of them expressed asTF (6/8). TF expression at the protein level did not correlate with the differentiation of the carcinoma cell line. All but two pancreatic cancer tissue samples stained positive for TF (17/19). In all samples of CP weak staining was restricted to pancreatic duct cells, whereas only a few subendothelial cells were positive in 9/20 of normal controls. TF and TAT levels in PCa patients were significantly elevated compared to controls whereas elevated F1 + 2 levels did not reach statistical significance compared to controls. In CP patients TAT and F1 + 2 levels proved to be significantly elevated compared to controls, although TAT elevation was less pronounced than in PCa patients.. We conclude that in addition to the upregulated expression of TF on the cell membrane, soluble TF might contribute to activation of the coagulation system in pancreatic cancer.

Topics: Adenocarcinoma; Aged; Blood Coagulation; Cell Line, Tumor; Female; Humans; Male; Middle Aged; Pancreatic Neoplasms; Thromboembolism; Thromboplastin

The case history is presented of a patient with Trousseau's syndrome in which tissue factor originating from lung cancer appeared responsible for recurrent DVT/PE. This is thought to be the first such case to be reported.

Topics: Adenocarcinoma; Adult; Humans; Lung Neoplasms; Male; Pulmonary Embolism; Recurrence; Syndrome; Thromboplastin; Venous Thrombosis

Pulmonary tumor thrombotic microangiopathy (PTTM) is a rare clinicopathological entity causing severe pulmonary hypertension. Its histological features include widespread tumor emboli along with fibrocellular intimal proliferation and thrombus formation in the small arteries and arterioles of the lungs. The result is occlusion or stenosis of the pulmonary vasculature, but the detailed pathogenesis has yet to be clarified in spite of the serious clinical manifestations. Herein is described the case of a 62-year-old man with a gastric adenocarcinoma who died of sudden cardiopulmonary arrest. The autopsy revealed advanced cancer disease as well as findings of PTTM, which seemed to be the cause of his unexpected death. The carcinoma cells were immunohistochemically positive for vascular endothelial growth factor (VEGF) and also for tissue factor (TF). There is another report suggesting that TF might play an important role in the pathogenesis of PTTM. Also, VEGF has been reported to be involved in a variety of forms of pulmonary hypertension and to be upregulated by TF. These findings suggest that VEGF and TF may be involved in the pathogenesis of PTTM. The present PTTM case, in which the tumor cells demonstrate the coexpression of VEGF and TF, is important in facilitating understanding of the lethal disorder in the future.

Topics: Adenocarcinoma; Humans; Immunohistochemistry; Lung; Lung Neoplasms; Male; Microcirculation; Middle Aged; Neoplastic Cells, Circulating; Stomach Neoplasms; Thromboembolism; Thromboplastin; Vascular Endothelial Growth Factor A

Tissue factor (TF), the primary physiologic initiator of coagulation cascade, is involved in the process of angiogenesis of various malignancies. This study was designed to investigate the correlation of TF expression to angiogenesis of gastric carcinoma, and to study its clinical significance.. The expression of TF and vascular endothelial growth factor (VEGF) in 80 specimens of gastric carcinoma and 20 specimens of normal gastric tissue was detected by EnVision immunohistochemistry. Tumor microvessel density (MVD) was evaluated using anti-CD34 antibody as an endothelial marker with the same technique as well.. Positive rates of TF and VEGF, and the mean value of MVD were significantly higher in gastric carcinoma than in normal gastric tissue (65.00% vs. 5.00%, P<0.01; 67.50% vs. 5.00%, P<0.05; and 36.14+/-9.94 vs. 12.10+/-3.27, P<0.05). TF expression was positively correlated with VEGF expression and MVD value (P<0.05). TF expression was correlated to the overall survival times of patients, TNM stage, and liver metastasis (P<0.05).. Overexpression of TF relates with angiogenesis and prognosis of gastric carcinoma.

Topics: Adenocarcinoma; Adult; Aged; Antigens, CD34; Female; Follow-Up Studies; Humans; Liver Neoplasms; Lymphatic Metastasis; Male; Microcirculation; Middle Aged; Neoplasm Staging; Neovascularization, Pathologic; Prognosis; Stomach Neoplasms; Survival Rate; Thromboplastin; Vascular Endothelial Growth Factor A

Tissue Factor (TF), the initiator of the extrinsic coagulation cascade, is overexpressed in a variety of cancers. TF is also expressed in normal human endometrium but little is known about its expression or regulation in endometrial cancer. We demonstrate herein that TF is expressed in the endometrial adenocarcinoma cell line Ishikawa. Furthermore, epidermal growth factor (EGF) induces a rapid and sustained increase in TF expression. Estradiol and progesterone had no effect on basal or EGF-induced TF expression in Ishikawa cells. In contrast to the pronounced and sustained upregulation at the protein level, EGF treatment elicited only a modest and transient increase in TF mRNA levels. This activity corresponded to the response observed from an exogenous TF promoter construct. However, the induction of TF was abrogated by cycloheximide as well as actinomycin-D, inhibitors or protein- and mRNA-synthesis, respectively, demonstrating that EGF mediates its effect through activation of the TF gene. Fractionation experiments showed that EGF increases TF presence in caveolin-I containing membrane fractions. Coagulation and invasion assays were used to explore the physiological implications of TF regulation. The results demonstrate that EGF-mediated induction of TF increases the procoagulant activity and invasive potential of Ishikawa cells. Furthermore, immunocytochemistry confirmed that TF is regulated by EGF in primary cultures of normal endometrial epithelial cells and malignant tumor cells. In conclusion, EGF-mediated upregulation of TF results in accumulation of this glycoprotein in caveolae-like membrane fractions and increased coagulative and invasive potential. Our results suggest that TF may play an integral role in endometrial carcinogenesis.

Topics: Adenocarcinoma; Anticoagulants; Blotting, Western; Cell Line, Tumor; Cells, Cultured; Coagulants; Cycloheximide; Dactinomycin; Detergents; Endometrial Neoplasms; Endometrium; Epidermal Growth Factor; Estradiol; Female; Gene Expression Regulation, Neoplastic; Glycoproteins; Humans; Immunohistochemistry; Nucleic Acid Synthesis Inhibitors; Octoxynol; Progesterone; Promoter Regions, Genetic; RNA, Messenger; Sucrose; Thromboplastin; Time Factors; Transfection; Ultracentrifugation; Up-Regulation

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

Blood coagulation is activated commonly in pancreatic carcinoma but the role of the tumor cell in this activation is undefined. Immunohistochemical procedures were applied to fixed sections of 22 cases of resected adenocarcinoma of the pancreas to determine the presence of components of coagulation and fibrinolysis pathways in situ. Tumor cell bodies stained for tissue factor: prothrombin: and factors VII, VIIIc, IX, X, XII, and subunit "a" of factor XIII. Fibrinogen existed throughout the tumor stroma, and tumor cells were surrounded by fibrin. Staining for tissue factor pathway inhibitor, and plasminogen activators was minimal and inconsistent. Plasminogen activator inhibitors -1, -2, and -3 were present in the tumor stroma, and on tumor cells and vascular endothelium. Extravascular coagulation activation exists associated with pancreatic carcinoma cells in situ that is apparently unopposed by naturally occurring inhibitors or the plasminogen activator-plasmin system. We postulate that such local coagulation activation may regulate growth of this malignancy. These findings provide a rationale for testing agents that modulate the blood coagulation/fibrinolytic system (that inhibit tumor growth in other settings) in pancreatic carcinoma.

Topics: Adenocarcinoma; Aged; Blood Coagulation Factors; Endothelium, Vascular; Female; Fibrin; Fibrinogen; Humans; Immunoenzyme Techniques; Male; Middle Aged; Neoplasm Proteins; Neoplastic Stem Cells; Pancreatic Neoplasms; Plasminogen Activator Inhibitor 1; Plasminogen Activator Inhibitor 2; Protein C; Protein S; Prothrombin; Stromal Cells; Thrombophilia; Thromboplastin

A 54-year-old man was diagnosed as having pancreatic cancer and disseminated intravascular coagulation. His plasma tissue factor level on the 11th hospital day was 996 pg/ml (normal range, 120-270 pg/ml). He was treated with gabexate mesilate, antithrombin III, and low-molecular-weight heparin. However, he died of multiple organ failure on the 17th hospital day. The histological finding was poorly differentiated ductal adenocarcinoma of the pancreas, and the production of tissue factor in this lesion was revealed. Tissue factor is a factor that initiates blood coagulation; thus, its expression in pancreatic cancer is one of the causes of coagulation abnormalities in this disease. Although one report has demonstrated immunoreactivity for tissue factor in pancreatic cancer, the patient's detailed clinical course was not mentioned in that report. This is the first report to prove that pancreatic cancer produced tissue factor in a patient with disseminated intravascular coagulation.

Topics: Adenocarcinoma; Disseminated Intravascular Coagulation; Humans; Male; Middle Aged; Pancreatic Neoplasms; Radiography; Thromboplastin

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

Vasculature development is thought to be an important aspect in the growth and metastasis of solid tumors. Among the angiogenic factors produced by tumor cells, vascular endothelial growth factor is considered to be the most potent and pathologically important. The synthesis of this growth factor has been shown to be modulated through Sp1 function following stimulation by tumor necrosis factor alpha (TNF-alpha). Oligodeoxynucleotides (ODNs) were synthesized with either the consensus sequence for Sp1 binding (Sp1 decoy ODNs) or a mutated form of this sequence (mt-Sp1 decoy ODNs). Using the hemagglutinating virus of Japan (HVJ)-liposome method, we transferred these ODNs into cultured cancer cells (A549 and U251 cells). The TNF-alpha-mediated expression of both VEGF and transforming growth factor beta1 and tissue factor (TF) by the cancer cells could be simultaneously suppressed to less than 30% by transfection of Sp1 decoy ODNs but not by mt-Sp1 decoy ODNs. In addition, in vitro invasiveness, synthesis of mRNA for urokinase-type plasminogen activator, and cell proliferation of both cell lines were also inhibited to 40% by the transfection of only Sp1 decoy ODNs. These results suggested that the Sp1 decoy strategy would be effective for regulating tumor growth by simultaneously reducing cancer cell (a) angiogenic growth factor expression, (b) proliferation, and (c) invasiveness.

Topics: Adenocarcinoma; Binding Sites; Cell Division; Cell Movement; Endothelial Growth Factors; Fluorescein-5-isothiocyanate; Fluorescent Dyes; Glioblastoma; Humans; Liposomes; Lung Neoplasms; Lymphokines; Neoplasm Invasiveness; Oligonucleotides; Respirovirus; RNA, Messenger; Sp1 Transcription Factor; Thromboplastin; Transcriptional Activation; Transfection; Transforming Growth Factor beta; Transforming Growth Factor beta1; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Tissue factor (TF), the physiological procoagulant, is expressed in pancreatic tissue as a result of malignant transformation. The aim of this investigation was to assess its role in pancreatic tumour cell invasion and primary tumour growth.. The full-length TF gene (1360 base pairs) was cloned into the plasmid DNA vector pcDNA3 in sense and antisense orientations, and these vectors were used to transfect the MIA PaCa-2 human pancreatic adenocarcinoma cell line. TF gene expression was characterized by Northern blot analysis, total cellular antigenic content by enzyme-linked immunosorbent assay and cell surface procoagulant activity by enzymatic assay. Invasion of tumour cells in vitro was determined by a standard Matrigel assay, and primary tumour growth was measured in immunodeficient mice.. Overexpression of the TF gene, confirmed by an increased signal on Northern blotting, was associated with increases in both total antigenic content for TF (P = 0.001) and cell surface procoagulant activity (P = 0.008) in sense cells compared with wild-type cells. Likewise, both in vitro tumour cell invasion (P = 0.001) and primary tumour growth (P = 0.007) were increased in sense transfectants.. Expression of TF enhances in vitro invasion and primary tumour growth of MIA PaCa-2 cells, suggesting that this procoagulant molecule might have a role in pancreatic tumour biology. Presented in part to the 83rd meeting of the Surgical Research Society, Oxford, UK, January 1996 and awarded the David Patey Prize, and in part to the 1997 Annual Meeting of the Association of Surgeons of Great Britain and Ireland, Bournemouth, UK, April 1997.

Topics: Adenocarcinoma; Animals; Blotting, Northern; Cell Division; Female; Humans; Mice; Neoplasm Invasiveness; Neoplasm Proteins; Neoplasm Transplantation; Pancreatic Neoplasms; RNA, Messenger; Thromboplastin; Transfection; Tumor Cells, Cultured

Expression of tissue factor (TF), a cellular initiator of the extrinsic coagulation cascade, is a feature of many malignant tumours and is intimately involved in the process of metastasis. SW-480 human colon adenocarcinoma cells responded to thrombin (1 U ml(-1)) or phorbol 12-myristate 13-acetate (PMA, 0.1 microM) with a 6.0-fold and a 7.7-fold increase in their procoagulant activity (PCA), respectively, after 4-6 h incubation in serum-free medium. The thrombin-enhanced PCA was significantly inhibited by complexing of thrombin with hirudin, or by serine protease inhibition with 3,4-dichloroisocoumarin. Both effects of thrombin and PMA on PCA in SW-480 cells were blocked by pretreatment of cells with cycloheximide or actinomycin D, indicating that the response required de novo protein and RNA synthesis. The thrombin-enhanced PCA depended on the activation of protein kinase C (PKC) as it was diminished by staurosporine and calphostin C. Moreover, stimulation of SW-480 cells by thrombin or PMA led to a significant increase in TF mRNA within 3 h as measured by the reverse-transcription PCR method, which was also dependent on the activation of PKC. The unaltered decay rate of thrombin-enhanced TF mRNA, evaluated after the addition of staurosporine, suggested that its inhibitory effect occurred at a transcription level. Our data suggest that thrombin enhances TF gene expression and protein synthesis in tumour cells in vitro via PKC activation. The induction of TF expression in tumour cells by thrombin indicates that tumour-associated PCA might have a positive-feedback effect on in vivo local propagation of thrombus by thrombin formation.

Topics: Adenocarcinoma; Coagulants; Colonic Neoplasms; Enzyme Activation; Enzyme Inhibitors; Hemostatics; Humans; Neoplasm Proteins; RNA, Messenger; Staurosporine; Tetradecanoylphorbol Acetate; Thrombin; Thromboplastin; Tumor Cells, Cultured

Several studies have previously demonstrated tissue factor (TF) expression in solid tumors. In our study, we evaluated by immunohistochemical staining TF expression in 79 cases of colorectal cancer and 17 cases of metastatic cancer of the liver from colorectal cancer, and investigated the relationship between the clinicopathological features and TF expression. TF was detected in the tumor of 57% of colorectal cancer patients, and its expression was significantly increased (p=0.01) in metastatic tumors (88%). TF expression was more commonly observed in metastatic tumors than in any Dukes' stage of primary cancer. In primary cancer, the detection of TF was more frequent in cases with lymph node metastasis (Dukes' C, 63%) or with hematogenous metastasis (Dukes' D, 82%) than in tumors without lymph node or hematogenous metastasis (Dukes' A and B, 46%, p=0.03). These results suggest that the expression of TF is related with the metastatic potential of colorectal cancer.

Topics: Adenocarcinoma; Adult; Aged; Biomarkers, Tumor; Cell Differentiation; Colorectal Neoplasms; Disease Progression; Female; Humans; Liver Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Proteins; Neoplastic Cells, Circulating; Prognosis; Thromboplastin

To investigate the expression of tissue factor (TF) and tissue factor pathway inhibitor (TFPI) in human colorectal carcinomas, Northern blot analysis was performed in a series of human colorectal carcinoma cell lines and in normal or tumoral colorectal tissues. Of 16 human colorectal carcinoma cell lines examined, most expressed TF mRNA, though the levels of expression varied significantly. Considerably higher expression was observed in the cell line CaR-1, while lines established from metastatic lesions tended to express abundant TF mRNA. By contrast, TFPI mRNA levels were low in these high TF-expressing cell lines. TFPI was expressed abundantly in WiDr and in a few other cell lines which expressed a very low level of TF mRNA. Immunocytochemically, both proteins were stained predominantly on the cell surface; however, diffuse cytoplasmic staining for TF also was observed in CaR-1 cells. In addition, the cell surface TF activity was significantly higher in CaR-1 cells than in WiDr cells, confirming the results of mRNA analysis. The level of TF mRNA in colorectal carcinoma tissue in vivo and its ratio to the normal counterpart also varied significantly among the cases. To search for a possible role of TF/TFPI in metastasis of colorectal carcinoma cells, the expression of these genes was compared between a rectal adenocarcinoma cell line, RCM-1, and its highly metastatic subline, RCM-1 L-10. Compared with the parent line, RCM-1 L-10 expressed 7.5-fold higher levels of TF mRNA, whereas TFPI expression was not altered significantly or even decreased slightly. The higher cellular TF activity was confirmed in the metastatic subline in comparison with the parent line.

Topics: Adenocarcinoma; Blotting, Northern; Colorectal Neoplasms; Humans; Immunohistochemistry; Lipoproteins; Liver Neoplasms; RNA, Messenger; Serine Proteinase Inhibitors; Thromboplastin; Tumor Cells, Cultured

Several tumour-derived factors have recently been identified which induce tissue factor (TF) expression in endothelial cells in vitro. However, there is only limited evidence that endothelial cells lining tumour blood vessels express elevated procoagulant activity (PCA) in vivo. We have investigated the effects of human breast and small cell lung cancer cell lines on the PCA of human micro- and macrovessel endothelial cell monolayers using a one-stage clotting assay, as well as detection of TF mRNA by RT-PCR. Only conditioned medium from the MDA-MB-231 breast adenocarcinoma cell line produced a consistent although transient increase in endothelial cell surface PCA, which was maximal by 6-9 hr. TF mRNA was detectable in the endothelial cells after 1 hr incubation with MDA-MB-231-conditioned medium and subsequently fell below detectable levels. Following 24 hr stimulation, nearly half the endothelial cell PCA was due to the presence of TF-containing membrane vesicles shed by the MDA-MB-231 cells. Consistent with these findings, the MDA-MB-231 cell line expressed high levels of cell surface-associated TF activity. Co-culture of MDA-MB-231 and endothelial cells for up to 5 days increased (approx. 118-fold) PCA associated with endothelial cell monolayers, due mainly to sequestration of shed tumour cell vesicles. Our results suggest that induction of TF de novo is not a common feature in the supporting endothelium of these tumour types.

Topics: Adenocarcinoma; Biological Factors; Blood Coagulation Tests; Breast; Breast Neoplasms; Capillaries; Carcinoma, Small Cell; Cells, Cultured; Coculture Techniques; Culture Media, Conditioned; Endothelium, Vascular; Female; Humans; Lung Neoplasms; Polymerase Chain Reaction; Thromboplastin; Tumor Cells, Cultured; Umbilical Veins

High-sensitivity thromboplastin reagents suitable for use in the prothrombin time (PT) assay are typically prepared from human brain and placenta, tissues that are in limited supply and subject to viral contamination. Cloning and expression of recombinant human tissue factor (TF) has enabled production of a new generation of thromboplastin reagents whose performance and utility are under active investigation. The purpose of this study was to determine the feasibility of producing a sensitive human thromboplastin reagent from a non-recombinant source: cultured human cells. Several human cell lines with apparently high constitutive TF synthesis were identified, and a viable thromboplastin reagent (Humaplastin) was produced from a human lung cell line via a non-conventional process that did not require reconstitution or rehydration of TF in cell membranes. When calibrated against BCT/253, a human brain international reference thromboplastin, Humaplastin exhibited a mean normal prothrombin time of 12.6 +/- 0.7 s (mean +/- SD: n = 20) and an International Sensitivity Index of 1.09 +/- 0.019. The performance of this reagent was well correlated (r = 0.983) with Thromborel S, a commercially available human placental thromboplastin reagent. Orthogonal least squares regression of the log PT values from the placental thromboplastin reagent versus Humaplastin and two recombinant TF-based thromboplastin reagents suggested that the latter three reagents are somewhat more sensitive than the placental thromboplastin reagent, although such differences should not be expected to have a significant impact on clinical utility. It is concluded that cultured human lung cells represent a suitable source of tissue thromboplastin for production of a high-sensitivity non-recombinant thromboplastin reagent.

Topics: Adenocarcinoma; Anticoagulants; Astrocytoma; Blood Coagulation Factors; Brain; Brain Neoplasms; Calibration; Carcinoma, Squamous Cell; Cells, Cultured; Choriocarcinoma; Feasibility Studies; Female; Glioblastoma; Histiocytosis, Langerhans-Cell; Humans; Indicators and Reagents; Lung; Lung Neoplasms; Neoplasm Proteins; Placenta; Prothrombin Time; Recombinant Proteins; Reference Standards; Sensitivity and Specificity; Thromboplastin; Tumor Cells, Cultured; Uterine Neoplasms

Procoagulant activity of pairs of cell lines, which were derived from the same original cell type but which possess different growth characteristics and metastatic properties, was examined. The following characteristics were considered suggestive of a greater likelihood of metastatic potential: high histological grade; establishment of the line from a metastatic rather than a nonmetastatic cancer; increased tumorigenicity in nude mice; and/or estrogen receptor-negative mammary cancer. Procoagulant activity was evaluated by a two stage clotting assay. Procoagulant activity was highly variable, with up to a 1,300-fold difference, among the cancer cell lines examined. The rate of clot formation was factor VII dependent and was totally inhibited by an anti tissue factor monoclonal antibody, indicating that tissue factor was the only significant procoagulant present in these cancer cells. Tissue factor antigen expression, evaluated by ELISA, correlated with procoagulant activity. In all pairs of cancer cell lines, those with characteristics of increased proliferative potential had increased tissue factor levels compared to cell lines that originated from the same cell type, but which possess less aggressive characteristics. Tissue factor activity in these cancer cells was increased by cell lysis or by exposure of intact cells to a calcium ionophore, similar to results previously obtained in fibroblasts. Tissue factor mRNA was evaluated by northern blot analysis using a specific probe complementary to tissue factor mRNA. The previously described predominant tissue factor mRNA species of 2.2 kb was identified in the majority of cancer cell lines examined, but tissue factor mRNA species of 3.2 to 3.4 kb were also identified.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Blood Coagulation; Blotting, Northern; Breast Neoplasms; Carcinoma, Renal Cell; Carcinoma, Transitional Cell; Cell Line; Gastrointestinal Neoplasms; Gene Expression; Humans; Kidney Neoplasms; Neoplasm Metastasis; RNA, Messenger; Thromboplastin; Tumor Cells, Cultured

We have studied the effects of two human pancreatic cancer and two human small cell lung cancer cell lines on clotting and platelet aggregation. Both pancreatic lines markedly shortened recalcification times and induced platelet aggregation. The lung cancer lines produced little shortening of recalcification times and no platelet aggregation. The clotting and aggregation activities of the pancreatic lines were further characterized. Recalcification times following the addition of cancer cell line material to plasmas deficient in factors VII and X were markedly prolonged, suggesting that the activity is due to tissue factor. Hirudin, an inhibitor of thrombin from the saliva of leeches, and rabbit polyclonal immunoglobulin G anti-bovine brain tissue factor inhibited both procoagulant and aggregation activities. Apyrase (an enzyme degrading ADP), diisopropylfluorophosphate (a serine protease inhibitor) and L-trans-epoxysuccinylleucylamido(4-guanidino)butane (a cysteine protease inhibitor) failed to inhibit these activities. Increasing concentrations of heparin inhibited platelet aggregation. Subcellular fractionation studies showed these activities to be localized to the plasma membrane. The association between mucin and the acceleration of clotting has been well described. The absence of mucin in electron micrographs of these pancreatic whole cells, membrane fractions, and shed microvesicles, as well as the failure of chaotropic agents (i.e., agents stripping material extrinsic to the cell membrane such as mucin) to abrogate this activity support these activities being intrinsic to the plasma membrane. These data strongly suggest that these activities are due to tissue factor which appears to be released as microvesicles in vitro. The release of tissue factor via microvesicles in vivo is one possible mechanism for the coagulopathy sometimes seen in patients with pancreatic carcinoma.

Topics: Adenocarcinoma; Blood Coagulation; Calcium; Carcinoma, Small Cell; Cell Membrane; Humans; Lung Neoplasms; Microscopy, Electron; Pancreatic Ducts; Pancreatic Neoplasms; Platelet Aggregation; Thromboplastin; Tumor Cells, Cultured

To analyze unique molecular differences between normal and neoplastic cells, we have examined host responses to tumor cells. The present study provides the first evidence for an innate rapid recognitive response of the lymphoid system to some syngeneic tumors. The lymphoid procoagulant (PCA) response, a T cell-instructed monocyte response that activates proteases of the coagulation cascade culminating in thrombin formation, is considered a component of classic delayed-type hypersensitivity responses. We have demonstrated that three syngeneic rat mammary carcinomas elicit this cellular response in vitro in lymphoid cells of the unimmunized rat. The response was rapid, reaching maximum within 6 hr. Analysis was compounded by the constitutive PCA activity of some tumors; however, the PCA product produced in the response to tumor challenge in vitro was newly biosynthesized and was of lymphoid cell origin, differing from the PCA of tumor cells. The lymphoid PCA response was prothrombinase-like and did not require vitamin K for biosynthesis, nor were other gamma-carboxylation-dependent extrinsic pathway proteases other than prothrombin required for thrombin generation. Both in vivo and in vitro derived mammary carcinoma cells elicited the response, whereas a fibrosarcoma and nontransformed syngeneic cells did not. Tumor shed substances, which were devoid of PCA and sedimentable only in part at 100,000 X G, induced this cellular response. The same stimuli shed from tumor cells did not directly elicit a PCA response from elicited peritoneal macrophages; however, in the presence of T lymphocytes a PCA response of these macrophages was produced. This study provides novel information to indicate that a T-enriched lymphocyte-dependent monocyte-macrophage response to some tumors, before effective in vivo immunization, may participate in initial local protease generation and fibrin deposition, both thought to play a significant role in the local pathobiology of tumors.

Topics: Adenocarcinoma; Animals; Cell Line; Culture Media; Female; Fibrosarcoma; Liver; Lymphocyte Activation; Lymphocytes; Macrophages; Mammary Glands, Animal; Mammary Neoplasms, Experimental; Monocytes; Rats; Rats, Inbred F344; Thromboplastin; Trypsin

Characterization studies have been carried out on 2 cell lines (KPK 1 and KPK 13) established from human renal adenocarcinoma. KPK 1 and KPK 13 have been passaged 178 times in vitro for about 6 years and 7 months and 78 times for about 3 years an 2 months, respectively. Although morphologic differences exist between the 2 lines, each has an epithelial morphology and exhibits multilayering. Doubling time of KPK 1 and KPK 13 cells was 29 hours and 51 hours, respectively. Both KPK 1 and KPK 13 induced tumors at the site of subcutaneous injection, closely resembling the original tumor from which they were derived. Chromosome number of both cell lines was 100 per cent aneuploid and the presence of Y chromosomes was confirmed by G banding in KPK 13 cells. KPK 1 was found to have high thromboplastic and high fibrinolytic activities, whereas KPK 13 was shown to have comparatively low thromboplastic and no detectable fibrinolytic activities. These activities were detected in the serum free supernatant fraction from KPK 1 cells but were not detected in that from KPK 13 cells.

Topics: Adenocarcinoma; Aged; Animals; Cell Line; Chromosome Banding; Female; Fibrinolysis; Humans; In Vitro Techniques; Karyotyping; Kidney Neoplasms; Mice; Mice, Nude; Neoplasm Transplantation; Thromboplastin

Analysis of fresh surgical specimens of normal tissue and tumor tissue show a cellular antithrombin activity to be present in certain organs. In normal tissues it was noted chiefly in normal colon, testes, breast, and uterus. In malignant tissues it was prominent in adenocarcinomas of the colon, breast, and lung. No epidermoid tumors showed evidence of thrombin binding. The thrombin- binding activity required the presence of intact cells and was distinct from the soluble antithrombins normally present in plasma and serum. There is growing evidence to suggest an interrelationship between clotting and the growth and dissemination of cancer. The implications of cellular antithrombins are reviewed in this context.

Topics: Adenocarcinoma; Antithrombins; Blood Coagulation Tests; Carcinoma, Squamous Cell; Female; Humans; Lymphoma; Male; Neoplasms; Thrombin Time; Thromboplastin

Topics: Adenocarcinoma; Adult; Aged; Animals; Blood Coagulation; Carcinoma; Disseminated Intravascular Coagulation; Female; Fibrinolysis; Humans; Male; Middle Aged; Prothrombin; Rabbits; Stomach Neoplasms; Thrombin; Thromboplastin

Topics: Abortion, Induced; Adenocarcinoma; Blood Coagulation Disorders; Carcinoma in Situ; Carcinoma, Squamous Cell; Curettage; Dilatation; Embryo, Mammalian; Female; Fetus; Gestational Age; Humans; Hypernatremia; Hypertonic Solutions; Lymphatic Metastasis; Oxytocin; Pregnancy; Prognosis; Thromboplastin; United States; Uterine Cervical Neoplasms; Uterine Neoplasms; Vaginal Smears

Topics: Adenocarcinoma; Adult; Aged; Alkaline Phosphatase; Alpha-Globulins; Biopsy; Fatty Liver; Female; Follow-Up Studies; Hepatitis; Humans; Kidney Neoplasms; Male; Middle Aged; Postoperative Care; Preoperative Care; Prothrombin Time; Serum Albumin; Sulfobromophthalein; Thromboplastin

Topics: Adenocarcinoma; Animals; Blood Platelets; Carcinoma 256, Walker; Cell Adhesion; Fibrin; In Vitro Techniques; Mammary Neoplasms, Experimental; Mice; Microscopy, Electron; Neoplasms, Experimental; Organoids; Platelet Adhesiveness; Pseudopodia; Rats; Thromboplastin

Topics: Adenocarcinoma; Blood Coagulation Tests; Carcinoma, Bronchogenic; Carcinoma, Squamous Cell; Coumarins; Female; Heparin; Humans; Male; Smoking; Thromboembolism; Thrombophlebitis; Thromboplastin