thromboplastin and Lung-Neoplasms

Inhibition of coagulation greatly limits cancer metastasis in many experimental models. Cancer cells trigger coagulation, through expression of tissue factor or P-selectin ligands that have correlated with worse prognosis in human clinical studies. Cancer cells also affect coagulation through expression of thrombin and release of microparticles that augment coagulation. In the cancer-bearing host, coagulation facilitates tumour progression through release of platelet granule contents, inhibition of Natural Killer cells and recruitment of macrophages. We are revisiting this literature in the light of recent studies in which treatment of clinical cohorts with anticoagulant drugs led to diminished metastasis.

Topics: Animals; Anticoagulants; Blood Coagulation; Blood Platelets; Cysteine Endopeptidases; Cytoplasmic Granules; Hirudins; Humans; Killer Cells, Natural; Lung Neoplasms; Macrophages; Mice; Neoplasm Metastasis; Neoplasm Proteins; Neoplasms, Experimental; Neoplastic Cells, Circulating; Neuraminidase; P-Selectin; Platelet Activation; Platelet Aggregation Inhibitors; Rats; Thrombin; Thrombophilia; Thromboplastin

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

Topics: Blood Coagulation; Gene Expression Regulation, Neoplastic; Lung Neoplasms; Neoplasm Metastasis; Thromboplastin

Topics: Animals; Blood Coagulation Disorders; Blood Platelets; Cytokines; Fibrinolysis; Humans; Lung Neoplasms; Macrophages; Thromboplastin

Coagulation activation and venous thromboembolism (VTE) are hallmarks of cancer; however, there is an unmet need of improved biomarkers for individualized anticoagulant treatment. The present sub-study of the RASTEN trial was designed to explore the role of coagulation biomarkers in predicting VTE risk and outcome in a homogenous cancer patient population. RASTEN is a multicenter, randomized phase-3 trial investigating the survival effect of low molecular weight heparin enoxaparin when added to standard treatment in newly diagnosed small cell lung cancer (SCLC) patients. Plasma collected at baseline, during treatment, and at follow-up was used in this ad hoc sub-study (N = 242). Systemic coagulation was assessed using four assays reflecting various facets of the coagulation system: Total tissue factor (TF); extracellular vesicle associated TF (EV-TF); procoagulant phospholipids (PPL); and thrombin generation (TG). We found small variations of biomarker levels between baseline, during treatment and at follow-up, and appeared independent on low molecular weight heparin treatment. Overall, none of the measured biomarkers at any time-point did significantly associate with VTE incidence, although increased total TF at baseline showed significant association in control patients not receiving low molecular weight heparin (P = 0.03). Increased TG-Peak was significantly associated with decreased overall survival (OS; P = 0.03), especially in patients with extensive disease. Low baseline EV-TF predicted a worse survival in the low molecular weight heparin as compared with the control group (HR 1.42; 95% CI 1.04-1.95; P = 0.03; P for interaction = 0.12). We conclude that the value of the analyzed coagulation biomarkers for the prediction of VTE risk was very limited in SCLC patients. The associations between TG-Peak and EV-TF with patient survival and response to low molecular weight heparin therapy, respectively, warrant further studies on the role of coagulation activation in SCLC aggressiveness.

Topics: Aged; Biomarkers, Tumor; Disease-Free Survival; Extracellular Vesicles; Female; Heparin, Low-Molecular-Weight; Humans; Incidence; Lung Neoplasms; Male; Middle Aged; Phospholipids; Small Cell Lung Carcinoma; Survival Rate; Thromboplastin; Venous Thromboembolism

Surgical treatment of lung cancer is associated with an elevated risk of thrombo-embolic complications. The question is whether the extent of pulmonary resection influences the concentration of serum coagulation system proteins.. This study aims to compare the blood coagulation activation parameters among patients undergoing pneumonectomy and lobectomy due to primary lung cancer.. A prospective study was carried out in 40 patients. Of whom, 30 underwent lobectomy and 10 treated with pneumonectomy. Serum concentrations of tissue factor (TF), tissue factor pathway inhibitor (TFPI), tissue factor pathway inhibitor-activated factor X complex (TFPI/Xa), thrombin-antithrombin complex (TAT), L-selectin, E-selectin and P-selectin were measured on the first and seventh postoperative days.. On the first postoperative day, the results of selected proteins concentrations were similar in both groups. However, on the seventh postoperative day, significantly higher concentrations of TF, TAT complex and E-selectin were found in patients who underwent pneumonectomy (median values: TF: 182.4 pg ml(-1) vs 116.6 pg ml(-1), P=0.031; TAT: 6.2 mg ml(-1) vs 3.9 mg ml(-1), P=0.048; E-selectin 40.24 ng ml(-1) vs 26.54 ng ml(-1), P=0.049).. Pneumonectomy was associated with significantly higher activation of coagulation system on the seventh postoperative day than lobectomy. TAT complex, TF and E-selectin are promising markers of extensive postoperative activation of coagulation and efficacy of antithrombotic prophylaxis.

Topics: Aged; Antithrombin III; Blood Coagulation; Carcinoma, Non-Small-Cell Lung; Female; Humans; Lipoproteins; Lung Neoplasms; Male; Middle Aged; Peptide Hydrolases; Pneumonectomy; Prospective Studies; Pulmonary Embolism; Selectins; Thromboplastin

Human blood monocytes (Mo) and monocyte-derived macrophages (M psi) possess cytotoxic effects against tumor cell lines when appropriately stimulated by various biological response modifiers, e.g., gamma interferon (gamma IFN) and muramyltripeptide (MTP). Activated Mo/M psi represent a new tool for the treatment of human malignancies, termed "adoptive cellular immunotherapy". Activated Mo/M psi express tissue factor procoagulant activity (PCA), which is a physiological trigger of blood coagulation. PCA was evaluated in vitro using a modification of the one-stage recalcification clotting time, and hemostatic changes were studied in vivo in cancer patients. Nine patients with peritoneal carcinomatosis were injected intraperitoneally with activated Mo and 11 patients with non-small cell lung carcinomas were infused intravenously with activated M psi. Hemostatic changes were followed using activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen level, antithrombin III (ATIII) and protein C (PC) activities. Fibrinolytic activity was estimated by euglobulin lysis time and assays for plasminogen and fibrin/fibrinogen degradation products (FDP). These assays were performed before and after each autologous infusion and on days 2 and 3. Activated Mo and M psi expressed potent PCA (85.5 +/- 7.5 U/ml for MTP activated Mo and 50 +/- 5.3 U/ml for gamma IFN activated M psi suspensions). In both groups of patients, APTT, PT, and TT underwent no significant variations. There was no significant consumption of ATIII or PC, and fibrinolysis was not activated during the study period. In the group injected intraperitoneally with MTP-activated Mo, fibrinogen showed a significant and progressive increase in relation to the development of an inflammatory reaction, reaching a maximum average value of 6.1 g/l at the end of the therapy with a concomitant increase in FDP levels. This increase was not observed after intravenous therapy with gamma IFN-activated M psi. No patient suffered from hemorrhagic or thrombotic events. In our experience, repeated injections of activated Mo or M psi expressing potent tissue factor PCA did not induce significant in vivo activation of the coagulation system in cancer patients.

Topics: Aged; Blood Coagulation Tests; Carcinoma, Non-Small-Cell Lung; Drug Evaluation; Female; Hemostasis; Humans; Immunotherapy, Adoptive; Lung Neoplasms; Macrophage Activation; Male; Middle Aged; Monocytes; Thromboplastin

Elevated tissue factor (TF) expression, although restricted in normal tissue, has been reported in multiple solid cancers, and expression has been associated with poor prognosis. This manuscript compares TF expression across various solid tumor types via immunohistochemistry in a single study, which has not been performed previously.. To increase insight in the prevalence and cellular localization of TF expression across solid cancer types, we performed a detailed and systematic analysis of TF expression in tumor tissue obtained from patients with ovarian, esophageal, bladder, cervical, endometrial, pancreatic, prostate, colon, breast, non-small cell lung cancer (NSCLC), head and neck squamous cell carcinoma (HNSCC), and glioblastoma. The spatial and temporal variation of TF expression was analyzed over time and upon disease progression in patient-matched biopsies taken at different timepoints. In addition, TF expression in patient-matched primary tumor and metastatic lesions was also analyzed.. TF expression was detected via immunohistochemistry (IHC) using a validated TF-specific antibody. TF was expressed in all cancer types tested, with highest prevalence in pancreatic cancer, cervical cancer, colon cancer, glioblastoma, HNSCC, and NSCLC, and lowest in breast cancer. Staining was predominantly membranous in pancreatic, cervical, and HNSCC, and cytoplasmic in glioblastoma and bladder cancer. In general, expression was consistent between biopsies obtained from the same patient over time, although variability was observed for individual patients. NSCLC biopsies of primary tumor and matched lymph node metastases showed no clear difference in TF expression overall, although individual patient changes were observed.. This study shows that TF is expressed across a broad range of solid cancer types, and expression is present upon tumor dissemination and over the course of treatment.

Topics: Carcinoma, Non-Small-Cell Lung; Female; Glioblastoma; Head and Neck Neoplasms; Humans; Lung Neoplasms; Male; Squamous Cell Carcinoma of Head and Neck; Thromboplastin

Thrombosis is a common complication of advanced cancer, yet the cellular mechanisms linking malignancy to thrombosis are poorly understood. The unfolded protein response (UPR) is an ER stress response associated with advanced cancers. A proteomic evaluation of plasma from patients with gastric and non-small cell lung cancer who were monitored prospectively for venous thromboembolism demonstrated increased levels of UPR-related markers in plasma of patients who developed clots compared with those who did not. Release of procoagulant activity into supernatants of gastric, lung, and pancreatic cancer cells was enhanced by UPR induction and blocked by antagonists of the UPR receptors inositol-requiring enzyme 1α (IRE1α) and protein kinase RNA-like endoplasmic reticulum kinase (PERK). Release of extracellular vesicles bearing tissue factor (EVTFs) from pancreatic cancer cells was inhibited by siRNA-mediated knockdown of IRE1α/XBP1 or PERK pathways. Induction of UPR did not increase tissue factor (TF) synthesis, but rather stimulated localization of TF to the cell surface. UPR-induced TF delivery to EVTFs was inhibited by ADP-ribosylation factor 1 knockdown or GBF1 antagonism, verifying the role of vesicular trafficking. Our findings show that UPR activation resulted in increased vesicular trafficking leading to release of prothrombotic EVTFs, thus providing a mechanistic link between ER stress and cancer-associated thrombosis.

Topics: Carcinoma, Non-Small-Cell Lung; Endoribonucleases; Guanine Nucleotide Exchange Factors; Humans; Lung Neoplasms; Pancreatic Neoplasms; Protein Serine-Threonine Kinases; Proteomics; Thromboplastin; Unfolded Protein Response

Small cell lung cancer (SCLC) patients have augmented risk of developing venous thromboembolism, but the mechanisms triggering this burden on the coagulation system remain to be understood. Recently, cell-derived microparticles carrying procoagulant phospholipids (PPL) and tissue factor (TF) in their membrane have attracted attention as possible contributors to the thrombogenic processes in cancers. The aims of this study were to assess the coagulation activity of platelet-poor plasma from 38 SCLC patients and to provide a detailed procoagulant profiling of small and large extracellular vesicles (EVs) isolated from these patients at the time of diagnosis, during and after treatment compared to 20 healthy controls. Hypercoagulability testing was performed by thrombin generation (TG), procoagulant phospholipid (PPL), TF activity, Protein C, FVIII activity and cell-free deoxyribonucleic acid (cfDNA), a surrogate measure for neutrophil extracellular traps (NETs). Our results revealed a coagulation activity that is significantly increased in the plasma of SCLC patients when compared to age-related healthy controls, but no substantial changes in coagulation activity during treatment and at follow-up. Although EVs in the patients revealed an increased PPL and TF activity compared with the controls, the TG profiles of EVs added to a standard plasma were similar for patients and controls. Finally, we found no differences in the coagulation profile of patients who developed VTE to those who did not, i.e. the tests could not predict VTE. In conclusion, we found that SCLC patients display an overall increased coagulation activity at time of diagnosis and during the disease, which may contribute to their higher risk of VTE.

Topics: Adult; Aged; Aged, 80 and over; Blood Coagulation Tests; Carcinoma, Small Cell; Centrifugation; Cysteine Endopeptidases; DNA; Extracellular Vesicles; Female; Humans; Lung Neoplasms; Male; Microscopy, Immunoelectron; Middle Aged; Nanoparticles; Neoplasm Proteins; Pulmonary Embolism; Risk Factors; Thrombin; Thrombophilia; Thromboplastin; Venous Thromboembolism

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

Within tumors, the coagulation-inducing protein tissue factor (TF), a major initiator of blood coagulation, has been shown to play a critical role in the hematogenous metastasis of tumors, due to its effects on tumor hypercoagulability and on the mediation of interactions between platelets and tumor cells. Targeting tumor-associated TF has therefore great therapeutic potential for antimetastasis therapy and preventing thrombotic complication in cancer patients. Herein, we reported a novel peptide-based nanoparticle that targets delivery and release of small interfering RNA (siRNA) into the tumor site to silence the expression of tumor-associated TF. We showed that suppression of TF expression in tumor cells blocks platelet adhesion surrounding tumor cells

Topics: Animals; Cell Line, Tumor; Female; Gene Expression Regulation, Neoplastic; Gene Silencing; Humans; Lung Neoplasms; Mice, Nude; Nanoparticles; Neoplasm Metastasis; Neoplasm Proteins; Neoplasms, Experimental; Neoplastic Cells, Circulating; RNA, Small Interfering; Thrombophilia; Thromboplastin; Thrombosis

To examine the expression of D-dimer, fibrinogen (FIB), leukocyte, C-reactive protein (CRP) and tissue factor (TF) released from monocyte in non-small cell lung cancer (NSCLC) patients with or without venous thromboembolism (VTE) and analyse the correlation, to explore the possible mechanisms.. Seventy-two patients confirmed the diagnosis of lung cancer, among whom 10 with VTE were enrolled into the study from November 2012 to January 2014 in the First Affiliated Hospital of Fujian Medical University and 30 healthy subjects were also enrolled as the control group. Ficoll and Percoll density gradient centrifugation separated of peripheral blood monocyte. Monocyte TF mRNA expression was detected using reverse transcriptase-polymerase chain reaction (RT-PCR).. There were significant differences in different stages of the cancer (P < .05) and no significance among the histopathologic types (P > .05) for the expression of monocyte TF mRNA in NSCLC patients, its expression was significantly higher in cancer with lymph node metastasis than those without lymph node metastasis (P < .01). Meanwhile, in NSCLC patients with VTE, the expression of monocyte TF mRNA was significantly higher than that in patients without VTE (P < .01). Difference of the survival curves between the low monocyte TF mRNA expression and the high monocyte TF mRNA expression was significant (Log-rank x2 = 4.923, P < .05).. Monocyte TF may be a relevant source of TF-mediated thrombogenicity in NSCLC patients and may be associated with prognosis in NSCLC.

Topics: Adult; Aged; Biopsy; Bronchoscopy; Carcinoma, Non-Small-Cell Lung; Female; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; Lymphatic Metastasis; Magnetic Resonance Imaging; Male; Middle Aged; Monocytes; Neoplasm Staging; Prognosis; Retrospective Studies; Reverse Transcriptase Polymerase Chain Reaction; RNA, Neoplasm; Thoracoscopy; Thromboplastin; Tomography, X-Ray Computed

Metastasis results from a complex set of traits acquired by tumor cells, distinct from those necessary for tumorigenesis. Here, we investigate the contribution of enhancer elements to the metastatic phenotype of osteosarcoma. Through epigenomic profiling, we identify substantial differences in enhancer activity between primary and metastatic human tumors and between near isogenic pairs of highly lung metastatic and nonmetastatic osteosarcoma cell lines. We term these regions metastatic variant enhancer loci (Met-VELs). Met-VELs drive coordinated waves of gene expression during metastatic colonization of the lung. Met-VELs cluster nonrandomly in the genome, indicating that activity of these enhancers and expression of their associated gene targets are positively selected. As evidence of this causal association, osteosarcoma lung metastasis is inhibited by global interruptions of Met-VEL-associated gene expression via pharmacologic BET inhibition, by knockdown of AP-1 transcription factors that occupy Met-VELs, and by knockdown or functional inhibition of individual genes activated by Met-VELs, such as that encoding coagulation factor III/tissue factor (F3). We further show that genetic deletion of a single Met-VEL at the F3 locus blocks metastatic cell outgrowth in the lung. These findings indicate that Met-VELs and the genes they regulate play a functional role in metastasis and may be suitable targets for antimetastatic therapies.

Topics: Carcinogenesis; Cell Line, Tumor; Enhancer Elements, Genetic; Epigenomics; Gene Expression Regulation, Neoplastic; Genome, Human; Humans; Lung Neoplasms; Neoplasm Metastasis; Osteosarcoma; Proteins; Selection, Genetic; Thromboplastin; Transcription Factor AP-1; Tumor Microenvironment

Platelets are thought to play an important role in metastasis formation, although the mechanisms involved remain incompletely understood. Here we studied the influence of platelet numbers on organ-specific metastasis to the lungs and lymph nodes using Tpo deficient mice that have low platelet counts. After tail vein injection of 4T1 breast cancer cells, the number of lung metastases was significantly lower in Tpo

Topics: Animals; Blood Platelets; Cell Line, Tumor; Female; Lung Neoplasms; Lymphatic Metastasis; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Thromboplastin

The procoagulant status of patients with non-small cell lung cancer (NSCLC) after chemotherapy is poorly characterized and the role of platelets in hypercoagulative state of NSCLC is unknown. The aim of this study was to evaluate the procoagulant activity (PCA) of platelets in NSCLC before and after chemotherapy. The subjects were 52 patients newly diagnosed with NSCLC. The patients had decreased clotting time compared with healthy subjects, and the thrombin-antithrombin complex increased 2.5-fold after chemotherapy. Platelets in the patients after chemotherapy had enhanced phosphatidylserine (PS) exposure, and shortened coagulation time as well as increased thrombin and fibrin formation of platelets compared with those before chemotherapy. Platelet-derived microparticles increased 2-fold at day 1 and peaked at day 2 post-chemotherapy. Treatment of cisplatin in vitro also resulted in upregulated intrinsic FXa and thrombin formation on platelets with a dose-dependent manner. Platelets treated with aspirin significantly decreased PCA. However, lactadherin blocked PS and inhibited the PCA approximately by 70%. Seven days after chemotherapy, PCA of platelets restored to the baseline as that before chemotherapy, indicating that within a week of chemotherapy patient platelets are highly procoagulant and effective intervention should be taken in case of thrombosis. Our results suggested that platelets after chemotherapy had elevated PCA and may contribute to the hypercoagulative state of NSCLC. Prophylactic anti-coagulant combined with anti-platelet therapy may play an inhibitory role in thrombotic complications in NSCLC.

Topics: Aged; Antigens, Surface; Aspirin; Bleeding Time; Blood Coagulation; Blood Platelets; Carcinoma, Non-Small-Cell Lung; Cell-Derived Microparticles; Female; Humans; Lung Neoplasms; Male; Middle Aged; Milk Proteins; Phosphatidylserines; Platelet Aggregation Inhibitors; Thrombin; Thromboplastin; Thrombosis; Up-Regulation

Zinc-binding protease aminopeptidase N (CD13) is expressed on tumor vascular cells and tumor cells. It represents a potential candidate for molecular targeted therapy, e.g. employing truncated tissue factor (tTF)-NGR, which can bind CD13 and thereby induce tumor vascular infarction. We performed a comprehensive analysis of CD13 expression in a clinically well characterized cohort of patients with small cell lung cancer (SCLC) to evaluate its potential use for targeted therapies in this disease.. CD13 expression was analyzed immunohistochemically in 27 SCLC patients and correlated with clinical course and outcome. In CD-1 nude mice bearing human HTB119 SCLC xenotransplants, the systemic effects of the CD13-targeting fusion protein tTF-NGR on tumor growth were tested.. In 52% of the investigated SCLC tissue samples, CD13 was expressed in tumor stroma cells, while the tumor cells were negative for CD13. No prognostic effect was found in the investigated SCLC study collective with regard to overall survival (p>0.05). In CD-1 nude mice, xenografts of CD13 negative HTB119 SCLC cells showed CD13 expression in the intratumoral vascular and perivascular cells, and the systemic application of CD13-targeted tissue factor tTF-NGR led to a significant reduction of tumor growth. We here present first data on the expression of CD13 in SCLC tumor samples. Our results strongly recommend the further investigation of tTF-NGR and other molecules targeted by NGR-peptides in SCLC patients. Considering the differential expression of CD13 in SCLC samples pre-therapeutic CD13 analysis is proposed for testing as investigational predictive biomarker for patient selection.

Topics: Aged; Animals; Biomarkers, Tumor; Blood Vessels; CD13 Antigens; Cell Line, Tumor; Female; Humans; Infarction; Kaplan-Meier Estimate; Lung Neoplasms; Male; Mice, Nude; Middle Aged; Molecular Targeted Therapy; Peptides; Recombinant Fusion Proteins; Small Cell Lung Carcinoma; Thromboplastin; Xenograft Model Antitumor Assays

Targeting cancer stem cell (CSC) represents a promising therapeutic approach as it can potentially fight cancer at its root. The challenge is to identify a surface therapeutic oncotarget on CSC. Tissue factor (TF) is known as a common yet specific surface target for cancer cells and tumor neovasculature in several solid cancers. However, it is unknown if TF is expressed by CSCs. Here we demonstrate that TF is constitutively expressed on CD133 positive (CD133+) or CD24-CD44+ CSCs isolated from human cancer cell lines, tumor xenografts from mice and breast tumor tissues from patients. TF-targeted agents, i.e., a factor VII (fVII)-conjugated photosensitizer (fVII-PS for targeted photodynamic therapy) and fVII-IgG1Fc (Immunoconjugate or ICON for immunotherapy), can eradicate CSC via the induction of apoptosis and necrosis and via antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity, respectively. In conclusion, these results demonstrate that TF is a novel surface therapeutic oncotarget for CSC, in addition to cancer cell TF and tumor angiogenic vascular endothelial TF. Moreover, this research highlights that TF-targeting therapeutics can effectively eradicate CSCs, without drug resistance, isolated from breast, lung and ovarian cancer with potential to translate into other most commonly diagnosed solid cancer, in which TF is also highly expressed.

Topics: A549 Cells; Animals; Antibody-Dependent Cell Cytotoxicity; Cell Line, Tumor; Female; Humans; Lung Neoplasms; Male; Metalloporphyrins; Mice; Mice, SCID; Molecular Targeted Therapy; Neoplastic Stem Cells; Ovarian Neoplasms; Photochemotherapy; Thromboplastin; Triple Negative Breast Neoplasms; Xenograft Model Antitumor Assays

Here we demonstrate a novel application of 'surface enhanced resonance Raman scattering nanoparticles' (SERRS NPs) for imaging breast cancer lung metastases with much higher precision than currently feasible. A breast cancer lung metastasis mouse model was established by intravenous injection of LM2 cells. These mice were intravenously administered SERRS NPs conjugated with ALT-836, an anti-tissue factor (TF) monoclonal antibody, and subjected to Raman imaging to visualize the expression of TF both in vivo and ex vivo. Raman imaging indicated marked uptake of αTF-SERRS-NPs by the lung metastases compared to isotype and blocking controls. Conversely, little uptake of αTF-SERRS-NPs was observed in the lungs of healthy mice. Successful detection and delineation of pulmonary micrometastatic lesions as small as 200 μm, corroborated by histology, immunohistochemistry, and bioluminescence imaging confirmed the suitability of both TF as a target and αTF-SERRS-NPs as an effective contrast agent for imaging breast cancer lung metastases. Further advancements of this technique in the form of Raman endoscopes coupled with ultrabright SERRS NPs developed in this work could lead to minimally invasive detection and resection of lung metastases.

Topics: Animals; Disease Models, Animal; Lung Neoplasms; Mice; Nanoparticles; Neoplasm Micrometastasis; Spectrum Analysis, Raman; 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

The initiator of extrinsic coagulation, tissue factor (TF), and its non-coagulant isoform alternatively spliced TF (asTF) are closely associated with tumor development. In the tumor microenvironment, the role of TF-induced coagulation in tumor progression remains to be fully elucidated. Using TF-knockdown lung tumor cells, we showed that TF is the dominant component of procoagulant activity but is dispensable in the cellular biology of tumor cells. In a xenograft model, using immunohistochemical analysis and flow cytometry analysis of the tumor microenvironment, we demonstrated that TF-induced fibrin deposition, which is correlated with complement activation and myeloid-derived suppressor cell (MDSC) recruitment, is positively associated with tumor progression. C5aR antagonism blunted the effect of TF on tumor progression and decreased MDSC recruitment. In conclusion, our data suggested that in tumor microenvironment, TF-induced coagulation activated the complement system and subsequently recruited myeloid-derived suppressor cells to promote tumor growth, which brings new insights into the coagulation-induced complement activation within the tumor microenvironment during tumor progression.

Topics: A549 Cells; Animals; Apoptosis; Blood Coagulation; Cell Proliferation; Complement Activation; Disease Progression; Female; Gene Knockdown Techniques; Humans; Lung Neoplasms; Mice, Nude; Myeloid-Derived Suppressor Cells; Receptor, Anaphylatoxin C5a; Thromboplastin

Epithelial-mesenchymal transition (EMT) is prominent in circulating tumor cells (CTC), but how it influences metastatic spread in this setting is obscure. Insofar as blood provides a specific microenvironment for tumor cells, we explored a potential link between EMT and coagulation that may provide EMT-positive CTCs with enhanced colonizing properties. Here we report that EMT induces tissue factor (TF), a major cell-associated initiator of coagulation and related procoagulant properties in the blood. TF blockade by antibody or shRNA diminished the procoagulant activity of EMT-positive cells, confirming a functional role for TF in these processes. Silencing the EMT transcription factor ZEB1 inhibited both EMT-associated TF expression and coagulant activity, further strengthening the link between EMT and coagulation. Accordingly, EMT-positive cells exhibited a higher persistance/survival in the lungs of mice colonized after intravenous injection, a feature diminished by TF or ZEB1 silencing. In tumor cells with limited metastatic capability, enforcing expression of the EMT transcription factor Snail increased TF, coagulant properties, and early metastasis. Clinically, we identified a subpopulation of CTC expressing vimentin and TF in the blood of metastatic breast cancer patients consistent with our observations. Overall, our findings define a novel EMT-TF regulatory axis that triggers local activation of coagulation pathways to support metastatic colonization of EMT-positive CTCs. Cancer Res; 76(14); 4270-82. ©2016 AACR.

Topics: Animals; Blood Coagulation; Cell Line, Tumor; Epithelial-Mesenchymal Transition; Female; Humans; Lung Neoplasms; Mice; Mice, Inbred BALB C; Neoplastic Cells, Circulating; Thromboplastin; Zinc Finger E-box-Binding Homeobox 1

Microvesicles (MV) in the blood stream are associated with distant metastasis in cancer. Platelet or endothelial cell-related MV actively participate in thrombogenesis, which is an important step in cancer metastasis. This study investigated the correlations between MV levels of platelet-poor plasma and distant metastasis in lung cancer.. Platelet-poor plasma from 44 treatment-naive lung cancer (23 with distant metastasis) and 19 normal subjects was used to determine the levels of glycoprotein Iβ (CD42) + platelet MV (PMV), P-selectin (CD62P) + PMV, VE-cadherin (CD144) + endothelial MV (EMV), tissue factor (CD142) + MV, thrombin-antithrombin complex and vascular endothelial growth factor (VEGF).. The level of CD142 + MV was significant (odds ratio 5.86, 95 % confidence interval 1.31-38.3) in predicting distant metastasis in lung cancer, and a cutoff value of 2.668 (after logarithm transformation) in the ROC curve had a specificity of 90 % and a sensitivity of 59 %. The presence of distant metastasis showed a significant correlation between CD144 + EMV and VEGF, but not between CD144 + EMV and CD42 + PMV or CD62P + PMV in lung cancer subjects.. The finding of CD142 + MV in platelet-poor plasma may be useful for suggesting distant metastasis in lung cancer. In addition to thrombogenesis, interaction between VE-cadherin and VEGF may be needed for successful metastasis in lung cancer.

Topics: Aged; Case-Control Studies; Cytoplasmic Vesicles; Female; Flow Cytometry; Humans; Lung Neoplasms; Male; Neoplasm Metastasis; ROC Curve; Thromboplastin

LKB1 encodes a serine/threonine kinase generally inactivated in many human cancers, which mediates cancer cell proliferation, migration and differentiation. Recent studies indicated that LKB1 exhibits potent anti-metastatic activity. However, the underlying molecular mechanisms of this activity remain unclear. In this study, we re‑introduced LKB1 into A549 lung cancer cells that lack the LKB1 gene to investigate how LKB1 affects tumor invasiveness and metastasis. We demonstrated that overexpression of the LKB1 protein in lung cancer cells resulted in significant inhibition of invasion. Furthermore, transfected lung cancer cells with LKB1 suppressed tissue factor (TF) and vascular endothelial growth factor (VEGF) expression at both the mRNA and protein levels. Here, we provided evidence showing that downregulation of TF and VEGF by LKB1 is correlated well with the inhibition of cell invasion. Overexpression of the LKB1 protein in human lung cancer is significantly associated with a decrease in activity and expression of the transcription factor SP1. Constitutive activation of the transcription factor Sp1 plays a critical role in TF and VEGF overexpression. We conclude that suppression of lung cancer cell invasion by LKB1 through downregulation of TF and VEGF may partly depend on its inhibitory effect on the transcription factor Sp1. Collectively, our data provide a novel molecular mechanism for the antitumor activity of LKB1 and may help further improve its effectiveness in controlling lung cancer growth and invasion.

Topics: AMP-Activated Protein Kinase Kinases; Cell Adhesion; Cell Line, Tumor; Cell Movement; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Humans; Lung Neoplasms; Protein Serine-Threonine Kinases; Sp1 Transcription Factor; Thromboplastin; Transfection; Vascular Endothelial Growth Factor A

Over expression of tissue factor (TF) occurs in more than 50 % of colorectal cancer (CRC). Therefore, TF represents an attractive target antigen for immunotherapy in CRC.. Here, we assessed the safety and efficacy of a recombinant adenovirus vector encoding the light chain of human coagulation factor VII (hfVII-LC) and human IgG1 Fc fragment (hIgG1-Fc), termed "benc vector," by targeting TF in the mouse model with colon cancer. Benc vector was administered intravenously or intratumorally in SCID mice with TF over-expressing HT-29 colon cancer. The safety and efficacy of benc vector were observed during animal experiments.. Complete inhibition of tumor growth (5/5) was observed not only in the intravenously injection of benc vector group but also in the intratumorally of benc vector group. We also observed a precautionary effect on lung metastases of HT-29 cells by intratumoral injection of benc vector. In the control group of animals given empty control vector, all animals (5/5) developed lung tumors and exhibited a higher number of nodules after injection with HT-29 cells via the tail vein. In contrast, only three animals (3/5) in the treatment group receiving benc vector had any observable lung metastases and a lower number of nodules. No animals died and no bleeding was observed both in treatment groups and control groups. Moreover, only moderate liver damage was detected in mice receiving benc vector by intravenous injections.. Benc vector encoding hfVII-LC and hIgG1-Fc can effectively inhibit tumor growth and metastases in SCID mice with TF over-expressing colon cancer and shows promise as an agent for CRC immunotherapy.

Topics: Adenoviridae; Animals; Colorectal Neoplasms; Factor VII; Genetic Vectors; HEK293 Cells; HT29 Cells; Humans; Immunoglobulin Fc Fragments; Immunoglobulin G; Immunohistochemistry; Immunotherapy; Injections, Intralesional; Injections, Intravenous; Lung Neoplasms; Mice; Mice, SCID; Recombinant Fusion Proteins; Thromboplastin; Treatment Outcome; Tumor Burden; Xenograft Model Antitumor Assays

Alternative splicing is a key regulatory mechanism for cellular metabolism controlling cell proliferation and angiogenesis, both of which are crucial processes for tumorigenesis under hypoxia. Human cells express two tissue factor (TF) isoforms, alternatively spliced TF (asTF) and 'full length' TF (flTF). flTF is the major source of thrombogenicity whereas, the function of soluble asTF, particularly in cancer, is widely unknown. In the present study, we examined the impact of alternative splicing on the pro-angiogenic potential and the TF expression pattern of A549 cells under hypoxia. We focused our efforts toward alternative splicing factors, such as Clk1, and pro-angiogenic proliferation-regulating factors, such as Cyr61. We further examined the influence of asTF overexpression on the expression of MCP-1, Cyr61 and VEGF, as well as on cell number and pro-angiogenic properties of A549 cells. Notably, we found hypoxia to induce the expression of alternative splicing factors (Clk1 and Clk4) as well as proliferation- and angiogenesis-promoting factors (Cyr61 and flTF). asTF overexpression in A549 cells also increased both cell number and tube formation. These effects were mediated by the induction of Cyr61, MCP-1 and VEGF, as well as by integrin α(v)β(3). Taken together, our results suggest that the pro-angiogenic potential of A549 lung cancer cells is modulated under hypoxic conditions via modulation of TF isoform expression which in turn is controlled by alternative splicing.

Topics: Alternative Splicing; Cell Hypoxia; Cell Line, Tumor; Cell Proliferation; Chemokine CCL2; Cysteine-Rich Protein 61; Humans; Integrin alphaVbeta3; Lung Neoplasms; Neovascularization, Pathologic; Protein Isoforms; Protein Serine-Threonine Kinases; Protein-Tyrosine Kinases; Thromboplastin; Vascular Endothelial Growth Factor A

Cytochalasin D (CytD) targets actin, a ubiquitous protein in eukaryotic cells. Previous studies have focused mainly on the antitumor effects of CytD. We previously found CytD to promote lung metastasis in B16 melanoma cells, which we had not anticipated, and, therefore, in the present study we investigated the possible underlying mechanisms. B16 melanoma cells were co-cultured with CytD and other agents and used to establish a lung metastatic model. In this B16 melanoma metastatic model, significantly increased lung metastasis and lung weight were found in CytD-treated mice, which was almost completely suppressed by tissue factor (TF) RNA interference expressed via lentivirus. The results of northern and western blot, and real-time RT-PCR analysis showed that the expression of TF was significantly upregulated in B16 cells treated with CytD but was significantly inhibited by TF RNA interference. In addition, upregulation and phosphorylation of mitogen-activated protein kinase p38 were also found in the metastatic lung tissues treated with CytD and in the B16 cells co-cultured with CytD and factor VIIa (FVIIa), but not in cells cultured with CytD, dimethyl sulfoxide or FVIIa alone. These results indicate that CytD stimulates the expression of TF in B16 melanoma cells, activating both coagulation-dependent and -independent pathways via binding to FVIIa, eventually promoting lung metastasis. TF interference is a potential approach to the prevention of B16 melanoma metastasis.

Topics: Animals; Cell Line, Tumor; Cytochalasin D; Factor VIIa; Female; Gene Expression Regulation, Neoplastic; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Nucleic Acid Synthesis Inhibitors; p38 Mitogen-Activated Protein Kinases; Phosphorylation; RNA Interference; RNA, Small Interfering; Thromboplastin; Up-Regulation

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

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

Tissue factor (TF), the transmembrane receptor for factor VIIa (FVIIa), has key regulatory functions in coagulation as well as in tumour progression and metastasis. Small-cell lung cancer (SCLC) metastasises more aggressively than non-small-cell lung cancer (NSCLC). Previously, we described the transition of SCLC cell line H69 to adherent growth and TF expression. Here, we explored the differential expression of TF and its functional impact on morphology and matrix metalloproteinase (MMP) secretion.. The constitutional TF expression was evaluated in a panel of established NSCLC and SCLC cell lines. Furthermore, in three stress-selected adherent SCLC H69 cells, TF and MMP expressions were determined by mRNA, protein, and activity measurements. RNA interference-mediated TF down-regulation and FVIIa stimulation were used to study the impact of TF on cellular functions.. NSCLC cells expressed high TF antigen (median 3.75 ng/mg; range 0.31-65.2 ng/mg protein, n = 8), while SCLC expressed none or low TF (median 0.07 ng/mg; range 0-0.39 ng/mg protein, n = 6). However, selected H69 adherent cells markedly expressed TF (range: 4.8-44.3 ng/mg protein, n = 3) and secreted MMP-2 and MMP-9. FVIIa stimulated MMP-2 and MMP-9 secretion in H69adh cells, whereas TF down-regulation diminished MMP-2 and MMP-9 expression and promoted reversion to suspension growth.. Our data show the significance of TF expression in the reversible growth phenotype of H69. Because TF, MMP expression, and adherence are highly relevant to cancer metastasis, this study suggests a novel mechanism of adaptation, thereby adding to the understanding of SCLC biology and its aggressiveness.

Topics: Cell Adhesion; Cell Line, Tumor; Cell Proliferation; Cell Survival; Factor VIIa; Fluorescent Antibody Technique; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Small Cell Lung Carcinoma; Thromboplastin

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

Tissue factor (TF), a rate-limiting enzyme cofactor in activating coagulation, is highly expressed in a wide spectrum of human tumor and tumor stromal cells. Using TF-deficient cancer cells and a conditional TF-knockout mouse model, we show that TF expressed by cancer cells, but not by the host stromal cells, plays a critical role in tumor growth. In the tumor microenvironment, serum coagulation factors are readily extravasated and therefore lead to continuous TF-mediated activation of coagulation proteases. To target this highly specific cascade of serine proteases, we used both a TF:VIIa inhibitor and doxorubicin-based prodrugs that are selectively activated by TF:FVIIa, FXa, and thrombin. Treatment with the TF:FVIIa inhibitor led to growth retardation in breast tumor models. In contrast, treatment with the prodrug eliminated primary tumor cells and lung metastases without apparent toxicity. Our findings offer preclinical proof of principle that targeting the coagulation cascade that is activated in the tumor microenvironment can be a highly effective approach for cancer therapy.

Topics: Animals; Blood Coagulation; Blood Coagulation Factors; Breast Neoplasms; Disease Progression; Doxorubicin; Factor VIIa; Female; Humans; Lung Neoplasms; Mammary Neoplasms, Animal; Mice; Mice, Knockout; Mice, Nude; Molecular Targeted Therapy; Prodrugs; Thrombin; Thromboplastin; Tumor Microenvironment

Cancer patients exhibit a high rate of thromboembolism (VTE). In this study, we analyzed levels of microparticle (MP) tissue factor (TF) activity in cancer patients with or without VTE. Blood was collected from cancer patients within 24 h of objectively diagnosed VTE (n=53) and from cancer patients without VTE (n=13). MPs were isolated from platelet poor plasma by centrifugation at 20,000g for 15 min. MP TF activity was measured using a two-stage chromogenic assay. Cancer patients with VTE had a significantly higher mean MP TF activity compared with cancer patients without VTE (1.7+/-3.8 pg/mL vs 0.6+/-0.4 pg/mL, p<0.05). Further prospective studies are required to determine if levels of MP TF activity may be a useful biomarker to identify patients at increased risk for VTE.

Topics: Biomarkers; Case-Control Studies; Cell-Derived Microparticles; Centrifugation; Chromogenic Compounds; Colonic Neoplasms; Humans; Lung Neoplasms; Neoplasms; Pancreatic Neoplasms; Thromboplastin; Venous Thromboembolism

Topics: Alternative Splicing; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Gene Expression Regulation, Neoplastic; Genetic Predisposition to Disease; Humans; Kaplan-Meier Estimate; Lung Neoplasms; Prognosis; Proportional Hazards Models; Risk Assessment; Risk Factors; RNA, Messenger; Thromboplastin; Time Factors; 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

It is well established that the blood coagulation system is activated in cancer. In addition, there is considerable evidence to suggest that clotting activation plays an important role in the biology of malignant tumors, including the process of blood-borne metastasis. For many years our laboratory has used experimental models of lung metastasis to study the events that follow the introduction of procoagulant-bearing tumor cells into circulating blood. This chapter focuses on the basic methods involved in assessing the anti-metastatic effects of anticoagulants and anti-platelet agents using rodent models of experimental metastasis. In addition, it summarizes our experience with these models, which collectively suggests that intravascular coagulation and platelet activation are a necessary prelude to lung tumor formation and that interruption of coagulation pathways or platelet aggregation may be an effective anti-metastatic strategy.

Topics: Animals; Anticoagulants; Antineoplastic Agents; Blood Coagulation; Cell Line, Tumor; Enoxaparin; Humans; Lung Neoplasms; Mice; Neoplasm Metastasis; Neoplasms, Experimental; Platelet Aggregation Inhibitors; Thromboplastin; Warfarin

Pulmonary tumor thrombotic microangiopathy (PTTM) is a rare clinicopathologic entity causing severe pulmonary hypertension, right-side heart failure, and sudden death. Its histologic features include widespread tumor emboli of the small arteries and arterioles of the lung, associated with thrombus formation and fibrocellular and fibromuscular intimal proliferation. The most frequent causative neoplasm for PTTM is gastric cancer, but lesions in other organs, including the ovary, have been occasionally identified as primary causes. Detailed molecular mechanisms underlying PTTM remain unclear, but some studies have suggested that tissue factor (TF) and vascular endothelial growth factor (VEGF) expressed by tumor cells may be involved in the pathogenesis for cases of gastric cancer. However, little is known about these molecules in PTTM caused by neoplasms of non-gastric origin. Here, we report the autopsy findings of a 42-year-old woman with ovarian cancer showing positive immunoreactivity for both TF and VEGF who died suddenly of PTTM. The present case provides support for the conclusion that these factors may be involved in the pathogenesis of PTTM, independent of the causal neoplasm.

Topics: Adenocarcinoma, Clear Cell; Adult; Antineoplastic Combined Chemotherapy Protocols; Autopsy; Chemotherapy, Adjuvant; Fatal Outcome; Female; Gynecologic Surgical Procedures; Heart Arrest; Humans; Immunohistochemistry; Lung Neoplasms; Neoplastic Cells, Circulating; Ovarian Neoplasms; Pulmonary Embolism; Thromboplastin; Thrombosis; Vascular Endothelial Growth Factor A

Cancer progression is facilitated by blood coagulation. Anticoagulants, such as Hirudin and low molecular weight heparins (LMWHs), reduce metastasis mainly by inhibition of thrombin formation and L- and P-selectin-mediated cell-cell adhesion. It is unknown whether the effects are dependent on cancer cell type. The effects of anticoagulants on tumor development of K1735 and B16 melanoma cells and CT26 colon cancer cells were investigated in mouse lung. Tumor load was determined noninvasively each week up to day 21 in all experiments using bioluminescence imaging. Effects of anticoagulants on tumor development of the three cell lines were correlated with the fibrin/fibrinogen content in the tumors, expression of tissue factor (TF), protease activated receptor (PAR)-1 and -4 and CD24, a ligand of L- and P-selectins. Hirudin inhibited tumor development of B16 cells in lungs completely but did not affect tumor growth of K1735 and CT26 cells. Low molecular weight heparin did not have an effect on K1735 melanoma tumor growth either. TF and PAR-4 expression was similar in the three cell lines. PAR-1 and CD24 were hardly expressed by K1735, whereas CT26 cells expressed low levels and B16 high levels of PAR-1 and CD24. Fibrin content of the tumors was not affected by LMWH. It is concluded that effects of anticoagulants are dependent on cancer cell type and are correlated with their CD24 and PAR-1 expression.

Topics: Animals; Anticoagulants; Blood Coagulation; CD24 Antigen; Cell Line, Tumor; Fibrin; Fibrinogen; Heparin, Low-Molecular-Weight; Hirudins; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; P-Selectin; Receptor, PAR-1; Thromboplastin; Transplantation, Heterologous

Recombinant nematode anticoagulant protein c2 (rNAPc2) is a specific inhibitor of tissue factor (TF)/factor VIIa complex with novel antithrombotic activity. TF is highly expressed in human colorectal tumors, and levels are positively correlated with disease progression.. To explore the therapeutic potential and mechanism of action of rNAPc2 during tumor growth and metastasis, we tested rNAPc2 in several experimental colorectal cancer models in mice.. Administration of rNAPc2 inhibited pulmonary metastasis in mice systemically disseminated with CT26 murine colon carcinoma cells in a dose-dependent fashion. Combining rNAPc2 with the cytotoxic agent 5-fluorouracil or bevacizumab (humanized anti-vascular endothelial growth factor monoclonal antibody) resulted in additive growth inhibition and simultaneous reduction of microvessel density in HCT116 human colorectal tumor xenografts in nude mice. Furthermore, rNAPc2 potentiated CPT-11 in inhibiting hepatic metastasis in nude mice with portal vein injection of HCT116 tumor cells. Long-term administration of rNAPc2 significantly suppressed spontaneous formation of intestinal tumors in Apc(Min/+) mice. Using a RNA interference approach, we showed that TF expression is necessary for rNAPc2-mediated inhibition of HCT116 human colorectal tumor xenograft growth in nude mice, indicating that the antitumor effect of rNAPc2 may be transduced through TF that is expressed on tumor cells.. rNAPc2 is a potent anticancer agent when used in combination with chemotherapy or antiangiogenic therapy in mouse models of colorectal cancer, and TF positivity appears to be required for its activity.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Bevacizumab; Cell Line, Tumor; Colorectal Neoplasms; Disease Progression; Female; Fluorouracil; Helminth Proteins; Humans; Intestinal Neoplasms; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Thromboplastin; Xenograft Model Antitumor Assays

Tissue factor (TF), the main initiator of blood coagulation, is also a signaling protein that regulates cancer progression. TF synthesis was recently shown to be affected by tumor suppressor genes (TSGs) in tumor cell lines. We therefore studied TF gene (F3) expression and the status of genes coding for tumor protein p53 (TP53), phosphatase and tensin homolog (PTEN), and serine/threonine kinase 11 (STK11) in non-small cell lung cancer (NSCLC). Heparanase (HPSE) gene expression was also measured because this endo-beta-D-glucuronidase was recently shown to enhance TF gene expression.. TF and heparanase mRNA expression was measured by real-time PCR in 53 NSCLC tumors. Exons 5-8 of TP53 were sequenced from genomic DNA. Mutations of PTEN and STK11 were screened by multiplex ligation-dependent probe amplification.. TF mRNA levels were significantly higher in T(3)-T(4) tumors (P = 0.04) and in stages III-IV of NSCLC (P = 0.03). Mutations of TP53, STK11, and PTEN were identified in 20 (37.7%), 21 (39%), and 20 (37.7%) of tumors, respectively. TF expression was higher in mutated TP53 (TP53(Mut)) (P = 0.02) and PTEN(Mut) (P = 0.03) samples. Moreover, TF mRNA increased from 2700 copies (no mutation) to 11 6415 when 3 TSG were mutated. Heparanase gene expression did not differ according to TF gene (F3) expression or TSG mutation. The median survival time was shorter in patients with tumor TF mRNA levels above median values (relative risk 2.2; P = 0.03, multivariate analysis) and when TP53 was mutated (relative risk 1.8; P = 0.02).. These results provide clear evidence that combined oncogene events affecting TSG dramatically increase TF gene expression in lung tumors. Moreover, this study suggests that TF gene expression could be used as a prognostic marker in NSCLC.

Topics: Adult; Aged; Aged, 80 and over; AMP-Activated Protein Kinase Kinases; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Glucuronidase; Humans; Kaplan-Meier Estimate; Lung Neoplasms; Male; Middle Aged; Mutation; Polymerase Chain Reaction; Proportional Hazards Models; Protein Serine-Threonine Kinases; PTEN Phosphohydrolase; Survival Rate; Thromboplastin; Tumor Suppressor Protein p53

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

Tissue factor (TF) is the physiological trigger of blood coagulation, but it could also have an important role in cancer by regulating VEGF expression and angiogenesis.. TF expression was studied by real-time PCR in lung tumors of 64 patients with non-small-cell lung cancer (NSCLC) and by immunohistochemical analysis. The gene expression of two VEGF isoforms, VEGF165 and VEGF189, was also evaluated. Microvascular density (MVD) was studied by measuring Von Willebrand Factor (VWF) mRNA levels and by immunohistochemistry using an anti-CD34 antibody.. TF mRNA levels were significantly lower than in corresponding non-affected lung tissues. However, TF expression was higher in T3-T4 tumors and this result was confirmed by immunohistochemistry. VEGF189 mRNA levels were ten times higher than those of VEGF165 and well correlated with TF mRNA levels. MVD was lower in the inner part of tumors than in the adjacent non-affected lung without being related to TF expression. Finally, codon 12 K-ras mutation was found in 8 lung carcinomas, and higher TF and VEGF189 mRNA levels were measured in mutated tissues (p < 0.001).. These results suggest that high TF expression in lung tumors may result from K-ras mutation and contribute to NSCLC progression, probably via mechanisms other than angiogenesis.

Topics: Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Cell Line, Tumor; Female; Gene Expression; Genes, ras; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Mutation; Neovascularization, Pathologic; Polymerase Chain Reaction; Reverse Transcriptase Polymerase Chain Reaction; Thromboplastin; Vascular Endothelial Growth Factor A; von Willebrand Factor

Topics: Alternative Splicing; Biomarkers, Tumor; Colonic Neoplasms; Gene Expression Regulation, Neoplastic; Humans; Liver Neoplasms; Lung Neoplasms; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sensitivity and Specificity; Thromboplastin; Up-Regulation

Thromboembolic complications are frequently associated with advanced cancer. Interestingly, one of the major initiators of blood coagulation, tissue factor (TF), is reported to be overexpressed in several tumor types and can be found on both tumor cells and tumor vasculature. Although the exact mechanisms have yet to be elucidated, TF expressed on tumor cells can trigger intracellular signaling events through various pathways that can lead to tumor angiogenesis, proliferation, and metastasis. There exists preclinical evidence that disruption of TF dependent signaling can effectively inhibit tumor cell migration, metastasis, and angiogenesis. Here, we report for the first time that an antibody to tissue factor can also prevent tumor growth in vivo. Prophylactic administration of CNTO 859, a humanized anti-human TF antibody, was shown to inhibit experimental lung metastasis of MDA-MB-231 human breast carcinoma cells by over 99% compared to a control antibody. Furthermore, therapeutic doses of CNTO 859 were shown to reduce tumor incidence and growth of orthotopically implanted MDA-MB-231 cells.

Topics: Animals; Antibodies, Monoclonal; Breast Neoplasms; Carcinoma; Cell Proliferation; Female; Humans; Immunoglobulin G; Lung Neoplasms; Mice; Mice, Inbred Strains; Thromboplastin; Xenograft Model Antitumor Assays

Trousseau's syndrome is a prothrombotic state associated with malignancy that is poorly understood pathophysiologically.. Here we report studies on the blood of a 55-year-old man with giant-cell lung carcinoma who developed a severe form of Trousseau's syndrome. His clinical course was dominated by an extremely hypercoagulable state. Despite receiving potent antithrombotic therapy, he suffered eleven major arterial and venous thrombotic events over a 5 month period. We examined the patient's blood for tissue factor (TF), the major initiator of coagulation, and found its concentration in his plasma to be forty-one-fold higher than the mean concentration derived from testing of 16 normal individuals.. Almost all of the TF in the patient's plasma was associated with cell-derived microvesicles, likely shed by the cancer cells.

Topics: Blood Coagulation; Carcinoma, Giant Cell; Cytoplasmic Vesicles; Enzyme-Linked Immunosorbent Assay; Factor VIIa; Humans; Immunohistochemistry; Lipoproteins; Lung Neoplasms; Lymph Nodes; Male; Middle Aged; Reference Values; Syndrome; Thromboplastin; Thrombosis

Pulmonary tumor thrombotic microangiopathy is an unusual malignancy-related respiratory complication characterized by multiple microthrombi and intimal myofibroblast proliferation. Its clinical manifestation is subacute respiratory failure with pulmonary hypertension. Herein is reported a case of pulmonary tumor thrombotic microangiopathy associated with gastric signet ring cell carcinoma. A 51-year-old woman with gastric cancer died of subacute respiratory failure. Autopsy showed gastric signet ring cell carcinoma with diffuse metastasis of pulmonary lymphatics and pleurae; every organ examined lacked a space-occupying tumor mass. Histologically, proliferated intimal myofibroblasts obliterated most of the pulmonary vascular lumen, and a few stenosed vascular lumina contained cancer cells. In addition, pulmonary vasculature associated with intimal proliferation contained microthrombi. Most cancer cells in the stomach and pulmonary lymphatics were typical signet ring cells, whereas those in vascular lesions were cells of poorly differentiated adenocarcinoma without mucous production. Consistent with a previous report, the latter expressed vascular endothelial growth factor (VEGF) and tissue factor (TF). The proliferated intimal myofibroblasts also expressed type 2A serotonin receptor (5-HT(2A)). These findings suggest that local expression of VEGF, TF, and 5-HT(2A) may be linked to the pathogenesis of this unusual pulmonary complication.

Topics: Carcinoma, Signet Ring Cell; Endothelium, Vascular; Fatal Outcome; Female; Humans; Hypertension, Pulmonary; Lung; Lung Neoplasms; Microcirculation; Middle Aged; Neoplastic Cells, Circulating; Receptor, Serotonin, 5-HT2A; Respiratory Insufficiency; Stomach Neoplasms; Thromboembolism; Thromboplastin; Vascular Endothelial Growth Factor A

We have shown that the thrombin G-protein coupled receptors (GPCR) designated as protease-activated receptors (PAR-1) are expressed in primary cancer cells isolated from peritoneal and pleural malignant effusions. Here, our main goal was to evaluate several coagulation and thrombin activation effectors and markers in a series of 136 malignant effusions from cancer patients with gastrointestinal, lung and mammary carcinomas. All these patients present a highly activated coagulation system in blood and their malignant effusions, as indicated by high levels of prothrombin F1.2 fragments and D-dimers. Notably, we detected in the effusions all the coagulation factors of the tissue factor pathway inducing thrombin activation, namely factors VII, V, X and II, as well as high VEGF levels and IGF-II in mature and precursor forms. Fibrin clot formation also correlated with higher levels of free ionized calcium (iCa), suggesting that iCa and its binding protein albumin are regulatory factors for fibrinogenesis in effusions. Consequently, thrombin, VEGF and IGFII appear to converge in the promotion of survival and invasivity of the metastatic cancer cells from blood to the malignant effusions. Thus, we add new insights on the interconnections between blood coagulation disorders in cancer patients and thrombin activation in malignant effusions, including their functional interaction with PAR in metastatic cancer cells. Based on these data we propose to counteract the metastatic cascades by targeted invalidation of key effectors of the coagulation system. Therefore, potential therapeutic approaches include the application of thrombin protease inhibitors, VEGF-blocking antibodies, and drugs targeting the VEGF and thrombin signaling pathways, such as tyrosine kinase or GPCR inhibitors.

Topics: Aged; Antineoplastic Agents; Antithrombins; Ascitic Fluid; Blood Coagulation; Blood Coagulation Factors; Breast Neoplasms; Calcium; Case-Control Studies; Factor V; Factor VII; Factor X; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Gastrointestinal Neoplasms; Humans; Insulin-Like Growth Factor II; Lung Neoplasms; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasms; Peptide Fragments; Pericardial Effusion; Pleural Effusion, Malignant; Proteins; Prothrombin; Serum Albumin; Thrombin; Thromboplastin; Vascular Endothelial Growth Factor A

Clotting activation and thromboembolic manifestations are common features in patients with cancer. Tumor cells can directly activate the clotting through two procoagulants: tissue factor (TF) and cancer procoagulant (CP).. The aim was to evaluate the levels of TF and CP in patients with different tumors in order to: (1) establish an association between these markers and the tumor localization, (2) establish a correlation between the levels of procoagulants and the status of the disease, (3) evaluate if the treatment with chemotherapy induced some modifications on the levels of procoagulants, (4) evaluate the possibility of using procoagulants as predictors in the development of thrombosis.. Sixty-one patients with different types of cancer (lung, breast, digestive and genitourinary) and 20 normal controls were included. The activity of TF and CP was studied in serum samples. Statistical analysis of the data was performed by two-tailed Fisher exact test.. The TF was increased in 72.5 and 0% (p < 0.01) of cancer patients and normal controls, respectively. PC was found to be increased in 88% of the cancer patients but in healthy controls it was increased in only 15% (p < 0.01). The patients with genitourinary cancer presented the highest values of both procoagulants coinciding with a major prevalence of thrombotic events. The activity CP was found in 93% of patients with stages I and II but in patients with stages II and IV disease it was found in 85% (not significant). There were no differences in the levels of both procoagulants between the patients treated with chemotherapy and those with other treatments.. TF and CP are elevated in patients with cancer. The highest values of both procoagulants are in the genitourinary cancer group in agreement with the greater presence of thrombosis observed in this group. Clinical follow up is important in order to determine the potential value of these procoagulants and the tendency to develop thrombosis in patients with cancer.

Topics: Adolescent; Adult; Breast Neoplasms; Case-Control Studies; Cysteine Endopeptidases; Digestive System Neoplasms; Disease Progression; Female; Humans; Lung Neoplasms; Male; Middle Aged; Neoplasm Proteins; Neoplasm Staging; Neoplasms; Predictive Value of Tests; Thromboplastin; Thrombosis; Urogenital Neoplasms

To evaluate the expression of tissue factor (TF), urokinase-type plasminogen activator (uPA), and urokinase-type plasminogen activator receptor (uPAR) in non-small cell lung cancer (NSCLC) tissues and to find their roles in lymph node metastasis, vascular involvement and prognosis.. Immunohistochemistry was used to examine the expression of TF, uPA, and uPAR in the tumor tissues of 97 NSCLC patients obtained during operation and 40 samples of normal lung tissues at least 5 cm away from the tumor tissues. The correlations of expression of TF, uPA, and uPAR with the clinicopathologic parameters were analyzed by chi2 test. The survival rates were calculated by Kaplan-Meier method.. TF, uPA, and uPAR were diffusely expressed in the carcinoma cell cytoplasm with the positive rates of 61.9%, 58.8%, and 61.9% respectively; however, they were only weakly expressed in the scattered macrophage and fibroblast cells in the normal lung tissues. TF expression was correlated with tumor angiogenesis as measured by microvessel density (P < 0.01); TF(34/47), uPA(33/47), and uPAR (39/47) expressions were all positively correlated with lymph node metastasis (P < 0.05, P < 0.05, and P < 0.01), and the uPAR expression was positively correlated with vascular involvement (P < 0.01). The agreement between TF and uPAR expression was significant (r = 0.432, P < 0.01). Co-expression of TF and uPAR was significantly correlated with lymph node metastasis and vascular involvement. Kaplan-Meier survival analysis showed that median the survival time of the patients with TF, uPAR and TF-uPAR positive tumor was shorter than that of the patients with TF, uPAR and TF-uPAR negative tumors (P < 0.01).. TF promotes angiogenesis, and uPAR contributes to lymph node metastasis and vascular involvement. Co-expression of TF and uPAR may play an important role in the metastasis and prognosis of NSCLC.

Topics: Adult; Aged; Carcinoma, Non-Small-Cell Lung; Female; Follow-Up Studies; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Lung; Lung Neoplasms; Lymphatic Metastasis; Middle Aged; Neoplasm Staging; Prognosis; Receptors, Cell Surface; Receptors, Urokinase Plasminogen Activator; Thromboplastin; Urokinase-Type Plasminogen Activator

The coagulation trigger tissue factor has been implicated in tumor growth, angiogenesis, and metastasis. In this study, we explore the effects of ex vivo and in vivo delivery of short interfering RNA (siRNA) targeting tissue factor on B16 melanoma colonization of the lung in a murine model for metastasis. The purposes of this work are to establish a noncytotoxic in vivo model for investigation of tissue factor function and provide preclinical assessment of the therapeutic potential of tissue factor siRNA for prevention of metastasis.. C57BL/6 mice were evaluated for pulmonary metastases following tail vein injection of B16 cells transfected with either active or inactive siRNA. Mice receiving cells transfected with active siRNA had significantly lower numbers of pulmonary tumors compared with mice injected with control cells (transfected with inactive siRNA). The average time point at which the mice started to exhibit tumor-associated stress was also increased significantly from 22 days for the control group to 27 days for the experimental group (P = 0.01). In a therapeutically more relevant model, where the siRNA was delivered i.p. and the cells (untransfected) by tail vein injection, an inhibitory effect on metastasis was observed when the siRNA treatment was initiated either before or at the time of cell injection.. The results suggest that tissue factor has a crucial function in promoting lung tumor metastasis of blood-borne tumor cells in the early stages of the tumor take process and further suggest that treatment with tissue factor siRNA may become a viable clinical strategy for prevention of tumor metastasis.

Topics: Animals; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Drug Delivery Systems; Female; Injections, Intravenous; Injections, Subcutaneous; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Transplantation; RNA, Small Interfering; 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

Topics: Blood; Carcinoma, Squamous Cell; Humans; Lung Neoplasms; Thromboplastin; Up-Regulation

Topics: Animals; Antibodies, Monoclonal; Baculoviridae; Blood Coagulation; Enzyme-Linked Immunosorbent Assay; Hybridomas; Immunohistochemistry; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasm Transplantation; Rats; Recombinant Proteins; Thromboplastin

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

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

To investigate the role of tissue factor(TF) in hematogenous metastasis of human colorectal carcinoma cells (LOVO) in vivo.. The eukaryotic expression vectors pcDNA3.1/Zeo bearing either sense or antisense TFc DNA were transfected into LOVO cells by lipofectamine 2000. TF protein expression in the transfected cells was detected by Western blot. Eighteen nude mice (Babl/c nu/nu) were randomly divided into three groups, and then transfected and untransfected LOVO cells were implanted via tail vein respectively. The nude mice were sacrificed 8 weeks after implantation, and the number of metastatic nodules in the lung was used to assess the metastatic ability of LOVO cells.. Compared with the untransfected group, TF expression of LOVO cells and the numbers of metastatic nodules in the lung increased in sense-TF cDNA transfection group (P< 0.05, P< 0.01, respectively), whereas decreased in antisense-TF cDNA transfection group (P< 0.05, P< 0.01, respectively).. TF can increase the hematogenous metastatic ability of human colorectal carcinoma cells (LOVO) in vivo.

Topics: Animals; Cell Line, Tumor; Colorectal Neoplasms; DNA, Complementary; Humans; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Invasiveness; Thromboplastin; Transfection

To investigate the role of tissue factor (TF) in the invasion and hematogenous metastasis of human colorectal carcinoma cells.. The eukaryotic expression vectors pcDNA3.1/Zeo bearing either sense or antisense TFcDNA were transfected into HT-29 and LoVo cells by the way of lipofactamine 2000. TF proteins in transfected cells were detected by Western Blot. Then the transfected and un-transfected tumor cells were implanted into nude mice (Balb/c Nu/Nu) to produce primary tumor, lung metastasis and liver metastasis respectively.. HT-29 and LoVo cells with sense-TFcDNA transfection showed increased TF expression compared with the cells without transfection, but the cells with antisense-TFcDNA transfection got the contrary change. The primary tumor growth and invasive range, lung metastasis and live metastasis all increased in sense transfectants but reduced in antisense transfectants.. TF can increase the invasion and hematogenous metastatic ability of human colorectal carcinoma cells.

Topics: Animals; Cell Line, Tumor; Colorectal Neoplasms; DNA, Antisense; DNA, Complementary; Humans; Liver Neoplasms; Lung Neoplasms; Male; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplastic Cells, Circulating; Thromboplastin; Transfection

Tissue factor (TF), an initiator of the extrinsic coagulation cascade, is also expressed in a wide range of cancer cells and plays an important role in cancer progression and metastasis, as well as in processes independent of the blood coagulation pathway. For example, by acting as an adhesion molecule enabling tissue invasion, TF may play a key role in the metastatic process and angiogenesis in non-small cell lung cancer (NSCLC).. To further investigate the role of TF on tumor cell invasion in NSCLC, we measured the TF mRNA expression in the tumors of 42 NSCLC patients using real-time quantitative reverse transcription - polymerase chain reaction carried out in a LightCycler. We then compared the TF mRNA expression with histological evidence of invasion of blood and lymphatic vessels by tumor cells.. Although there was no significant relationship between the TF mRNA expression and the invasion of lymphatic vessels, the TF mRNA expression was significantly higher in tumors that invaded blood vessels (Log(10) TF mRNA/GAPDH mRNA = 2.16 +/- 0.18) than in those that did not (1.59 +/- 0.16; P = 0.03).. These results suggest that TF plays a major role in blood vessel invasion by tumor cells in NSCLC.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Neovascularization, Pathologic; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; RNA, Neoplasm; Thromboplastin

Tissue factor (TF) is the membrane receptor of the serine protease coagulation factor VIIa (FVIIa). Formation of the TF/FVIIa complex initiates the coagulation cascade. We used short hairpin RNA (shRNA)-mediated RNA interference to knock down TF expression in the human metastatic melanoma cell line LOX-L. After transfection with the shRNA construct, 3 stable clones with significantly downregulated TF expression were established. They exhibited decreased proliferation in vitro as determined by (14)C thymidine incorporation and soft agar assay. The in vivo metastatic potential was assessed in an experimental pulmonary metastasis model in which cells from different clones were injected into the tail vein of nude mice. The incidence of pulmonary tumors was significantly lower in mice receiving shRNA-expressing cells (33% +/- 15%) than in control mice injected with wild-type cells or cells stably transfected with empty expression vector (90% +/- 10%). The mice injected with TF-downregulated cells had markedly longer survival time (69 +/- 17 days) compared to the control mice (35.6 +/- 5 days; p = 0.03). Thus, reduction of TF levels in LOX-L cells significantly delayed and reduced lung tumor formation. As a first step in elucidating the molecular basis for this effect, we compared the global gene expression profile in TF-downregulated cells and control cells by using cDNA microarray analysis. Forty-four known human genes were found to be significantly upregulated (> 2-fold; p < 0.05) and 228 genes significantly downregulated (>or= 3-fold; p < 0.05) in TF-downregulated cells compared to control cells. The differentially expressed genes encode proteins functioning in transcription, translation, cell communication and cell growth/death. The results provide a basis for investigating molecular mechanisms underlying the effects of TF on the metastatic capacity of LOX-L melanoma cells.

Topics: Animals; Cell Line, Tumor; DNA, Neoplasm; Down-Regulation; Gene Expression Regulation, Neoplastic; Humans; Incidence; Lung Neoplasms; Melanoma; Mice; Mice, Nude; Oligonucleotide Array Sequence Analysis; RNA, Neoplasm; Serine Endopeptidases; Survival Rate; Thromboplastin; Transfection

An association between cancer and thrombosis has been recognized for more than a century. However, the manner by which tumor growth is regulated by coagulation in vivo remains unclear. To assess the role of coagulation on tumor growth, in vivo, we tested coagulation inhibitors specific for either tissue factor (TF)/factor VIIa (fVIIa) complexes or factor Xa (fXa) for antitumor activity. Here, we show that two inhibitors of TF/fVIIa, TF pathway inhibitor (TFPI) and the nematode anticoagulant protein rNAPc2, inhibit both primary and metastatic tumor growth in mice. In addition, we show that rNAPc2 is also a potent inhibitor of angiogenesis. In contrast, rNAP5, a second nematode anticoagulant protein that specifically inhibits fXa, does not exhibit antitumor activity. Because the hemostatic activity of TF/fVIIa is mediated through activation of fXa, these data suggest that proteolytic activity of TF/fVIIa promotes tumor growth and angiogenesis through a novel proangiogenic mechanism and independently of hemostasis.

Topics: Animals; Anticoagulants; Cell Division; Factor VIIa; Helminth Proteins; Lipoproteins; Lung Neoplasms; Male; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neovascularization, Pathologic; Thromboplastin

Tissue factor, the cellular initiator of blood coagulation, has been implicated as a determinant of metastatic potential in human melanoma cells. Here, we report that differential expression of tissue factor in murine melanoma cell lines of known metastatic behavior is mediated by AP-1-dependent and 12S E1A oncoprotein-repressible gene transcription. When compared to weakly metastatic C10 cells, highly metastatic M4 cells possessed elevated levels of tissue factor cofactor activity, transfected promoter activity, and heterodimeric AP-1 DNA-binding complexes containing Fra-1. Transient co-expression of the adenovirus E1A 12S oncoprotein strongly repressed transcription of an AP-1-driven tissue factor reporter gene indicating the additional requirement of N-terminal E1A-interacting coactivators. Stable expression of E1A mutants defective in CBP/p300-binding failed to suppress tissue factor expression and experimental metastasis by M4 cells while clones expressing wild type E1A exhibited greatly reduced tissue factor cofactor activity and metastatic potential in vivo. Overexpression of functional tissue factor in cells containing wild type E1A failed to restore the highly metastatic M4 phenotype suggesting that additional E1A-responsive and CBP/p300-dependent genes are required to facilitate metastasis of murine melanoma cells demonstrating high tissue factor expression and cofactor activity.

Topics: Adenovirus E1A Proteins; Animals; E1A-Associated p300 Protein; Gene Expression Regulation, Neoplastic; Hematologic Neoplasms; Lung Neoplasms; Melanoma; Mice; Mice, SCID; Models, Theoretical; Neoplasm Metastasis; Nuclear Proteins; Promoter Regions, Genetic; Protein Structure, Tertiary; Proto-Oncogene Proteins c-fos; Thromboplastin; Trans-Activators; Transcription Factor AP-1; Transcription, Genetic; Tumor Cells, Cultured

Small cell lung cancer (SCLC) is a very malignant tumor known to grow aggressively and to metastasize early. It is well established that metastasis generally involves both tumor cell adhesion and proteolytic degradation of the extracellular matrix. However, SCLC cells cultured in vitro, such as the classic SCLC cell line NCI-H69, grow in floating aggregates and express only negligible proteolytic activity. In this report, we show that NCI-H69 cells can be selected for adherent growth. In contrast to parental suspension cells, the adherent cells were found to express tissue factor as well as gelatinolytic activity, attributable to matrix metalloproteinases 2 and 9. Such a switch of tumor cell characteristics, if it could occur in SCLC patients, might add to the understanding of the steps involved in the spreading of this highly metastatic type of lung cancer.

Topics: Carcinoma, Small Cell; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; Matrix Metalloproteinases; Thromboplastin; Tumor Cells, Cultured

From April 1986 to September 2000, 122 MRCC patients were treated by monthly intralymphatic injections (containing a mean of 573 IL-2 U and 26 x 10(6) LAK cells) and i.m. administration of IFN and TF; 71 patients also received a 3-day cycle of monthly IL-2 inhalations with a mean of 998 daily U. MRCC cases not treated by immunotherapy (n = 89) represent our historical controls. Adverse clinical side effects related to treatment were negligible. CR (n = 11) and PR (n = 13) were noticed in 24/122 patients. Of 24 responding patients, 17 resumed progression, whereas 7 remain in remission 11-69 months later. The overall median survival of treated patients (28 months) was 3.5-fold higher than the median survival of historical controls (7.5 months), and a Kaplan-Meier curve showed 25% survival 11 years after the beginning of immunotherapy. Apparently, the addition of IL-2 by inhalation improved survival. The present immunotherapy protocol appears to be efficacious, safe, devoid of adverse side effects, far less costly than others and able to offer a good quality of life to MRCC patients; if confirmed in a multicenter trial, it could set the basis for developing low-dose immunomodulatory treatments.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Female; Humans; Immunotherapy, Adoptive; Interferons; Interleukin-2; Kidney Neoplasms; Killer Cells, Natural; Lung Neoplasms; Male; Middle Aged; Thromboplastin

Tissue factor (TF) is a transmembrane glycoprotein that complexes with factor VIIa to initiate blood coagulation. We previously reported that expression of high levels of TF in a human melanoma cell line promotes metastasis. Both the cytoplasmic domain of TF and its extracellular domain complexed with factor VIIa are required for the metastatic effect. To further explore the mechanism of TF-mediated metastasis, we investigated the possibility that a protease-activated receptor (PAR) might play a role. For this purpose, we first determined the expression levels of the known PARs (PAR1-4) in a human melanoma cell line, SIT1, that has low endogenous levels of TF and low metastatic potential. We found negligible levels of all of the known PARs and transfection of this cell line with human TF cDNA did not alter expression of the known PARs. To study the possible role of PAR1 in TF-mediated metastasis, we prepared a panel of transfected cell lines with varying levels of TF and PAR1. Our studies show that TF promotes metastasis by a pathway that does not involve high expression of known PARs by tumor cells. In addition, while overexpression of PAR1 is insufficient to induce metastasis in cells with low TF expression, it enhances the metastatic potential of cells with high TF expression, indicating a possible synergy between TF and PAR1 in promoting metastasis.

Topics: Animals; Disease Models, Animal; Drug Synergism; Female; Humans; Lung Neoplasms; Melanoma; Mice; Mice, SCID; Neoplasm Metastasis; Receptor, PAR-1; Receptors, Thrombin; Thromboplastin; Transfection; Tumor Cells, Cultured

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) is an initiator of the extrinsic cascade of blood coagulation. Although recent studies have revealed a relationship between metastatic properties and TF expression in some neoplastic cells, the significance of TF in lung cancer, especially in non-small-cell lung cancer (NSCLC), is still unclear. In this study, TF was detected in NSCLC cell lines by functional study, Western blot analysis and immunocytochemical staining. TF levels in eight NSCLC cell lines were also quantitated by enzyme-linked immunosorbent assay (ELISA), and TF expression was evaluated in 55 specimens of surgically resected NSCLCs. NSCLC cell lines derived from metastatic lesions produced high levels of TF (48.3+/-23.5 ng 10(-6) cells, mean +/- s.e.m.), whereas those derived from primary lesions produced low levels of TF (0.2+/-0.1 ng 10(-6) cells). Immunohistochemical studies disclosed significantly stronger staining for TF in cells from NSCLC patients with metastasis than in those without metastasis. Among the 28 patients with metastasis, ten were strongly positive, 16 were moderately positive and two were negative for TF. In contrast, among the 27 patients without metastasis, only two were strongly positive, 18 were moderately positive and seven were negative for TF. Therefore, malignant cells from patients with lung cancer produce various levels of TF, and TF may play an important role in the metastatic process.

Topics: Biomarkers, Tumor; Blotting, Western; Carcinoma, Non-Small-Cell Lung; Enzyme-Linked Immunosorbent Assay; Humans; Immunohistochemistry; Lung Neoplasms; Neoplasm Metastasis; Thromboplastin; Tumor Cells, Cultured

Tissue factor (TF) is the physiological initiator of blood coagulation. It has been suggested that TF also regulates tumor growth and angiogenesis. We therefore used immunohistochemistry to analyze the expression of TF and angiogenesis in non-small-cell lung carcinomas of 191 patients. A significant association was found between TF expression and microvessel density (MVD): TF-negative carcinomas more frequently exhibited low MVD. Additionally, a significant relationship between TF expression and the expression of vascular endothelial growth factor (VEGF) was discovered. TF was also compared with the resistance of the carcinomas to doxorubicin, as measured in vitro: TF-negative tumors were more frequently resistant to doxorubicin than were TF-positive tumors. Kaplan-Meier survival analysis revealed that survival times were longer in patients with TF-negative tumors than in patients with TF-positive tumors. These data suggest that TF functions as an additional angiogenic factor that could be used as a new prognostic and predictive factor for non-small-cell lung carcinomas.

Topics: Carcinoma, Non-Small-Cell Lung; Humans; Immunohistochemistry; Lung Neoplasms; Middle Aged; Neovascularization, Pathologic; Prognosis; Survival Analysis; Thromboplastin

Tissue factor (TF), the initiating cell surface receptor of the coagulation cascade, plays important roles in embryogenesis, angiogenesis, and tumor cell metastasis. It is controversial whether proteolytic function of TF complexed with its serine protease ligand VIIa is required for metastatic tumor dissemination. We show here in a model for TF-dependent experimental hematogenous metastasis, that TF supports metastasis by both proteolytic activity of the TF-VIIa complex and currently undefined functions of the cytoplasmic domain. We demonstrate that ligand binding of VIIa to TF is required for metastasis. Antimetastatic properties of covalently inactivated VIIa provide evidence that ligand binding is insufficient per se to support metastasis, emphasizing that proteolytic activity is necessary for the metastatic process. Ala or Asp mutations of cytoplasmic serine residues were introduced to preclude or mimic phosphorylation. In vivo analysis of these mutants suggests that local protease generation on the tumor cell surface does not serve simply to activate the cytoplasmic domain of TF by serine phosphorylation. Thus, extracellular functions of the catalytically active TF-VIIa complex cooperate with specific functions of the TF cytoplasmic domain to support the complex process of hematogenous tumor cell dissemination.

Topics: Animals; CHO Cells; Cricetinae; Cytoplasm; Factor VIIa; Female; Lung Neoplasms; Mice; Mice, SCID; Neoplasm Metastasis; Phosphorylation; Structure-Activity Relationship; Thromboplastin; Transfection

Mononuclear cells (MNC) generate cell-bound procoagulant activity (PCA) which shortens recalcification time after incubation with an antigen to which the donor has been sensitized. PCA has been demonstrated in various lung diseases, including exudative pleural effusions. To determine the value of measuring cell-bound PCA in the diagnosis of tuberculous pleural effusions we examined pleural effusion MNC of patients with tuberculosis (n = 19), congestive heart failure (n = 7), and carcinoma (n = 7). MNC were isolated, incubated in 0 or 10 micrograms/ml purified protein derivative (PPD) for 15 min and for 20 h, and recalcification time determined. Incubation with thromboplastin was used as control. The recalcification times in serum incubated for 15 min varied within a wide range, the mean values were longest for tuberculous effusion MNC, incubation for 20 h increased variation. Incubation of cells for 15 min with thromboplastin led to a decrease of mean recalcification time in tuberculous (p < 0.001) and heart failure (p < 0.05), and with no significance in carcinomatous effusions. Incubation with PPD led to decrease of recalcification time which was not significant. Comparisons of the mean relative recalcification times after PPD incubation showed that tuberculosis differed from lung cancer (p < 0.001), lung cancer from heart failure (p < 0.05), but not heart failure from tuberculosis. We conclude from our study that pleural effusion MNC express spontaneous PCA in vitro which is strongest in carcinomatous pleural effusions. Incubation of MNC with thromboplastin and less discernable with PPD leads to an increase in PCA which is more pronounced in tuberculous pleural effusions. However, due to substantial intersubject variability and overlap between the study groups, this test does not allow reliable differentiation of tuberculous from other MNC rich pleural effusions.

Topics: Blood Coagulation; Blood Coagulation Tests; Cell Count; Diagnosis, Differential; Female; Heart Failure; Humans; Leukocytes, Mononuclear; Lung Neoplasms; Male; Middle Aged; Pleural Effusion; Pleural Effusion, Malignant; Thromboplastin; Tuberculin; Tuberculosis, Pleural

Although abnormalities of alveolar fibrin turnover have been reported to play a role in the development of idiopathic pulmonary fibrosis (IPF), the pathophysiological relevance remains unclear. We therefore investigated the localization of tissue factor (TF) and fibrin deposition in patients with IPF using immunohistochemistry and compared the results with those from patients who had interstitial pneumonia associated with systemic sclerosis (IP-SSc) and idiopathic bronchiolitis obliterans with organizing pneumonia (BOOP). Expression of TF-mRNA was also assessed, using in situ hybridization with a digoxigenin-labeled cRNA probe. In patients with IPF, IP-SSc, and idiopathic BOOP, the TF antigen was positively stained in type II pneumocytes and in some alveolar macrophages. The fibrin antigen was stained in the type II pneumocytes and the adjacent area. Tissue factor-mRNA was expressed in the type II pneumocytes and in some alveolar macrophages. Neither TF antigens nor TF-mRNA were detected in the normal lung. These results indicate that type II pneumocytes are a major source of TF, suggesting that TF production in these cells is closely related to fibrin deposition in the lungs of people with these diseases.

Topics: Base Sequence; Biopsy; Cryptogenic Organizing Pneumonia; DNA Primers; Fibrin; Humans; Immunohistochemistry; In Situ Hybridization; Lung; Lung Neoplasms; Molecular Sequence Data; Polymerase Chain Reaction; Pulmonary Fibrosis; RNA, Messenger; Scleroderma, Systemic; Thromboplastin

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

The human melanoma cell line M24met expresses tissue factor, the cellular initiator of the blood coagulation cascade. Blocking of the coagulation pathways at the level of tissue factor, factor Xa, or thrombin inhibits hematogenous M24met metastasis in SCID mice, implicating a role for thrombin generation in this process. Dependent on cell surface tissue factor activity, M24met cells generate thrombin in vitro. Thrombin and the thrombin receptor agonist peptide TRP-14 activate a signaling pathway in M24met cells that involves an increase in intracellular calcium and induces cell proliferation. Immunofluorescence evidences expression of the signaling thrombin receptor on these cells. Thus, M24met melanoma cells express both the initiating cell surface receptor for the coagulation pathways and the central signaling receptor of the coagulation system, suggesting the in situ generation of proliferative signals which can contribute to the malignant phenotype.

Topics: Animals; Antibodies, Monoclonal; Calcium; Cell Division; Cell Line; Cytosol; Female; Flow Cytometry; Fluorescent Antibody Technique; Humans; Kinetics; Lung Neoplasms; Melanoma; Mice; Mice, SCID; Peptide Fragments; Receptors, Thrombin; Signal Transduction; Thrombin; Thromboplastin; Transplantation, Heterologous; Tumor Cells, Cultured

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

Metastasis is a multistep process which requires highly adapted interactions of tumor cells with host target organs. Compared with nonmetastatic cells, metastatic human melanoma cells express 1000-fold higher level of tissue factor (TF), the major cellular initiator of the plasma coagulation protease cascades. To explore whether TF may contribute to metastatic tumor dissemination, we analyzed the effect of specific inhibition of TF function on human melanoma metastasis in severe combined immunodeficient (SCID) mice. Using species-specific antibodies to TF, we demonstrate that initial adherence in insufficient for successful tumor cell implantation in a target organ. Rapid arrest of human tumor cells in the lungs of mice was not diminished by inhibition of TF. However, inhibition of TF receptor function and consequent reduction in local protease generation abolished prolonged adherence of tumor cells, resulting in significantly reduced numbers of tumor cells retained in the vasculature of the lungs. The growth of pulmonary metastases was also significantly inhibited by a blocking anti-TF monoclonal antibody and Fab fragments thereof, whereas a noninhibitory antibody lacked antimetastatic effects. Cell surface expression of functional TF thus contributes to melanoma progression by allowing metastatic cells to provide requisite signals for prolonged adhesive interactions and/or transmigration of tumor cells across the endothelium, resulting in successful metastatic tumor implantation.

Topics: Animals; Cell Adhesion; Female; Gene Expression; Humans; Lung Neoplasms; Melanoma; Mice; Mice, SCID; Neoplasm Metastasis; Neoplasm Transplantation; RNA, Messenger; RNA, Neoplasm; Thromboplastin

Peri-tumour fibrin is a consistent feature of tumour stroma and is deposited shortly after tumour cell inoculation. Since there are several ways in which fibrin may be beneficial to tumour growth, it is possible that the ability of normal or malignant tissue to generate fibrin may influence metastasis. Many normal tissues and tumour cells possess a procoagulant activity that is due to a complex of tissue factor and factor VII. We have measured this tissue procoagulant activity in normal rats, rats stabilised on Warfarin and similarly anticoagulated animals injected with factor VII. The effect of Warfarin and factor VII administration on pulmonary seeding following injection of MC28 fibrosarcoma cells was also assessed. Procoagulant activity in adrenal, lung and colon was significantly reduced by Warfarin (P less than 0.001). Administration of factor VII significantly increased lung and adrenal tissue procoagulant activity in anticoagulated rats (P less than 0.02). Warfarinised rats had significantly slower primary tumour growth (P less than 0.001) and fewer lung deposits than control animals (P less than 0.001). Injection of factor VII restored pulmonary seeding to control levels (P less than 0.001). Warfarin did not affect the ability of the cells to adhere in vitro and did not reduce the number of tumour cells physically trapped in the lungs after intravenous injection. It is concluded that the procoagulant activity of normal tissues may influence their ability to support tumour growth and that the antimetastatic effect of Warfarin may be at least partly due to a reduction in the availability of the factor VII required for this activity.

Topics: Adrenal Glands; Animals; Cell Adhesion; Colon; Factor VII; Factor X; Fibrosarcoma; Liver; Lung; Lung Neoplasms; Neoplasm Seeding; Neoplasms, Experimental; Rats; Thromboplastin; Warfarin

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

Cultured SK-OS-10 cells (human osteogenic sarcoma metastatic to lung) shed microvesicles (dia. 300-1000 nm) that contained procoagulant and proaggregatory activities inhibitable by hirudin, by anti-tissue factor antibody and by phospholipase A2. These results show that SK-OS-10 cells belong to a group including U87MG human glioblastoma and HL-60 promyelocytic leukemia in which these activities are due to a thrombin-dependent mechanism arising from the presence of tissue factor on the surface of the tumor cells and their shed microvesicles.

Topics: Antibodies; Blood Platelets; Cell Line; Creatine Kinase; Hirudins; Humans; Inclusion Bodies; Lung Neoplasms; Osteosarcoma; Phosphocreatine; Phospholipases; Platelet Activating Factor; Platelet Aggregation; Thromboplastin

The mechanisms by which B16, 3LL and MH134 tumor cells induce platelet aggregation were studied. The B16 and 3LL tumors, which have high or moderate procoagulant activities, aggregated platelets only in the presence of Ca2+ and plasma factors. MH134, which had much lower procoagulant activity, aggregated platelets even in the absence of these factors. The induction of aggregation by B16 and 3LL could be prevented by thrombin inhibitors but not by the ADP scavenger, suggesting that thrombin, generated by procoagulant activities of tumor cells themselves, might play a major role in initiating aggregation. MH134-induced aggregation was not affected by any of the inhibitors, indicating that the mechanisms by which MH134 initiate platelet aggregation are independent of both thrombin and ADP.

Topics: Adenosine Diphosphate; Animals; Blood Coagulation; Calcium; Liver Neoplasms, Experimental; Lung Neoplasms; Male; Melanoma; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Neoplasm Metastasis; Platelet Aggregation; Thrombin; Thromboplastin

Fibrin was detected by specific immunofluorescence in tissue obtained from five of six cases of small cell carcinoma of the lung. Dense specific fluorescence was observed in the connective tissue stroma surrounding metastatic tumor nodules and frequently in the scant extracellular stroma surrounding individual viable tumor cells and small tumor cell clusters. When observed by electron microscopy, the fibrin hugged tumor cell plasma membranes and, in some areas, seemed to envelop the cells. Fluorescent staining of tumor cells, but not the stroma, was observed with an antibody to tissue factor. These findings suggest that local activation of coagulation occurs in small cell carcinoma of the lung. Deposited fibrin may contribute to the growth and spread of this particular type of cancer.

Topics: Antigens; Carcinoma, Small Cell; Fibrin; Fluorescent Antibody Technique; Humans; Lung Neoplasms; Microscopy, Electron; Thromboplastin

Three cultured cell lines of human lung cancer with varied degrees of thromboplastic and fibrinolytic activities were injected subcutaneously or intravenously into congenitally athymice nude mice, and their fate was observed with light and electron microscopes. All the cell lines formed solid tumor masses at the sites of subcutaneous injection. When the cancer cells with high or moderate thromboplastic activity were injected intravenously, marked platelet aggregation and fibrin deposition, followed by neutrophilic infiltration, were seen around the cancer cells arrested in pulmonary arterioles or capillaries. Platelet aggregation and fibrin deposition were less conspicuous, when the cancer cells with low thromboplastic activity were injected. No metastatic foci were found 2 weeks after the intravenous injection of each cancer cell line. These results suggest that thromboplastic activity of human cancer cells is closely related to thrombus formation at the sites of lodgement and that thrombus develops without participation of immunological mechanisms.

Topics: Animals; DiGeorge Syndrome; Fibrin; Fibrinolysis; Humans; In Vitro Techniques; Lung Neoplasms; Male; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplasm Transplantation; Neoplasms, Experimental; Platelet Aggregation; Thromboplastin; Transplantation, Heterologous

Crude tumor cell extract from rat ascites hepatoma AH130 revealed a high thromboplastic activity. Intravenous injection of the extract caused widespread thrombus formation in the capillaries, arterioles, and arteries of the lung. Intravenous inoculation of AH130 after an injection of the tumor cell extract produced metastatic foci in the larger arteries, compared with the case injected with only AH130 which developed metastatic foci mainly in and around the alveolar septa. These results suggest the role of tumor thromboplastin material in the mode of distribution of metastatic foci in the lung.

Topics: Animals; Liver Neoplasms, Experimental; Lung Neoplasms; Male; Pulmonary Artery; Pulmonary Embolism; Rats; Thromboplastin

Thromboplastic and fibrinolytic activities of 14 lines of cultured human cancer cells were estimated by modified Astrup's methods. High tissue thromboplastic activity was found in one line of urinary-bladder cancer, 2 lines of gastric cancer and one line of lung cancer, but no activity was found in 6 lines of lung cancer. High fibrinolytic activity was noted in one line of gastric cancer and 2 lines of lung cancer, but no activity was seen in 6 lines of lung cancer and one line of gastric cancer. No plasmin activity was found. The tumour cell lines could be classified into 3 groups on the basis of the 2 activities. Cancer cell lines could also be classified into 2 groups: with high or low release of thromboplastin into culture media. Fibrinolytic activity was found in the culture media of all cell lines with high fibrinolytic activity. Fibrinolytic activity, but not thromboplastic activity, seemed to be influenced by the constituents of culture media. No definite correlation was found between the 2 activities and the histological types of the parent tumours of the cultured cells.

Topics: Blood Coagulation; Cell Line; Factor IX; Factor VII; Fibrinolysis; Humans; Lung Neoplasms; Neoplasms; Stomach Neoplasms; Thromboplastin; Urinary Bladder Neoplasms

Thromboplastic and fibrinolytic activities of rat ascites tumor cells avoiding any stromal elements were examined and the role of these activities in the blood-borne metastasis was discussed. Ten lines of tumor cells showed varied thromboplastic and fibrinolytic activities. Tumor cell lines examined were classified into four groups; (1) lines AH-130, AH-62, and AH-7974 with high thromboplastic and high fibrinolytic activities, (2) lines AH-130F(N), AH-66F, and AH-7974F with low thromboplastic and low fibrinolytic activities, (3) line SLC with high thromboplastic and low fibrinolytic activities, and (4) lines AH-109A, AH-41A, and AH-41C with moderate thromboplastic and low fibrinolytic activities. The cell lines AH-130 and AH-130F(N), as well as AH-7974 and AH-7974F, have the same origin and showed different enzymic activities. AH-130 caused more prominent thrombus formation in the pulmonary vessels of rats in the early stage of intravenous inoculation and induced more prominent decrease in the number of platelets and fibrinogen levels in peripheral blood than AH-130F(N). Also, AH-130 developed more abundant metastatic foci in the lung 72 hr and 7 days after intravenous inoculation than AH-130F(N).

Topics: Animals; Blood Coagulation; Cell Line; Fibrinolysis; Liver Neoplasms, Experimental; Lung Neoplasms; Neoplasm Metastasis; Neoplasms, Experimental; Plasminogen Activators; Pulmonary Artery; Pulmonary Embolism; Rats; Thromboplastin

Topics: Adult; Aged; Animals; Blood Coagulation Disorders; Carcinoma, Bronchogenic; Carcinoma, Ehrlich Tumor; Disseminated Intravascular Coagulation; Female; Fibrin Fibrinogen Degradation Products; Fibrinogen; Fibrinolysin; Fibrinolysis; Heparin; Humans; Lung Neoplasms; Male; Mice; Middle Aged; Neoplasm Metastasis; Neoplasms; Neoplastic Cells, Circulating; Thromboembolism; Thromboplastin

Topics: Aged; Blood Cell Count; Blood Coagulation Disorders; Blood Coagulation Tests; Blood Platelets; Disseminated Intravascular Coagulation; Female; Fibrinogen; Fibrinolysin; Fibrinolysis; Hemoptysis; Humans; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Plasminogen; Prothrombin Time; Thrombin; Thrombophlebitis; Thromboplastin

Topics: Blood Coagulation Disorders; Humans; Lung Neoplasms; Methods; Pneumonectomy; Thromboplastin

Topics: Breast Neoplasms; Female; Humans; Lung Neoplasms; Middle Aged; Necrosis; Neoplasm Metastasis; Neoplasm Seeding; Neoplasms, Radiation-Induced; Radiotherapy; Skin Neoplasms; Thromboplastin

Topics: Blood Coagulation Factors; Erythrocyte Count; Factor V; Factor VII; Factor X; Fibrinogen; Fibrinolysis; Gastrointestinal Diseases; Humans; Kidney Diseases; Liver Diseases; Lung Neoplasms; Lymph; Prothrombin; Prothrombin Time; Thoracic Duct; Thromboplastin