fibrin and Lung-Neoplasms

To review derangements of pathways of fibrin turnover that promote pathologic fibrin deposition in the acute respiratory distress syndrome and to review the contribution of the endothelium and parenchymal lung cells to the derangements. In addition, to review how these pathways can be exploited in specific clinical circumstances, including sepsis and acute lung injury. Lastly, to review newly recognized posttranscriptional and urokinase-dependent pathways by which the fibrinolytic system is regulated in the lung.. Medical literature published in English from 1966 to present.. Local abnormalities of fibrin turnover in the injured lung recapitulate the systemic changes observed in sepsis. In both circumstances, the procoagulant response is increased, whereas fibrinolytic activity is concurrently depressed. The increased procoagulant activity is related to tissue factor associated with factor VII/VIIa. Fibrinolytic activity in the vasculature is mainly attributable to tissue plasminogen activator, whereas extravascular fibrinolytic activity in the lung is mainly attributable to urokinase plasminogen activator (uPA). Depressed fibrinolytic activity is in large part attributable to plasminogen activator inhibitor-1. In sepsis, activated protein C is also deficient, potentiating the inflammatory response, coagulopathy, and depressed fibrinolysis. Recombinant human activated protein C (drotrecogin alfa [activated]) was successful as an intervention for sepsis in a recent phase 3 clinical trial (PROWESS). Recently, novel posttranscriptional pathways that regulate expression of uPA, its receptor (uPAR), and plasminogen activator inhibitor-1 have been identified. The responsible mechanisms involve cis-trans interactions between newly recognized messenger RNA (mRNA) binding sequences and mRNA binding proteins. A 51 nucleotide mRNA binding sequence within the coding region of uPAR mRNA interacts with a novel 50-kDa mRNA binding protein to destabilize the message. Sequences within the 3' untranslated region of uPA or plasminogen activator inhibitor-1 mRNA interact with 30- and 60-kDa proteins, respectively, to regulate message stability. All of these pathways operate in lung epithelial cells, and endothelial cells regulate uPA expression through a similar pathway. In addition, uPA itself is capable of inducing expression of other components of the fibrinolytic system, including uPAR. This observation defines another feedback loop that could amplify local fibrinolysis and other uPA- or uPAR-mediated cellular responses, including cellular proteolysis, proliferation, and directed cellular migration.. Novel posttranscriptional pathways regulate expression of uPA, uPAR, and plasminogen activator inhibitor-1. uPA itself is capable of inducing other components of the fibrinolytic system. Some or all of these newly recognized pathways are operative in endothelial and parenchymal lung cells and may influence disordered fibrin turnover in the injured lung.

Topics: Endothelium, Vascular; Fibrin; Fibrinolysis; Humans; Lung Neoplasms; Plasminogen Activator Inhibitor 1; Respiratory Distress Syndrome; Urokinase-Type Plasminogen Activator

Topics: Fibrin; Humans; Lung; Lung Neoplasms; Pleura; Pneumonia

Lysyl oxidase (LO) initiates the first step in the crosslinking of collagens and elastin and has also been shown to function as a tumor suppressor. The purpose of the present work was to determine whether the products of a newly described LO-like gene (LOXL) that encodes a close homolog of LO, the LO-like (LOL) protein, is associated with extracellular matrix remodeling during fibrotic disorders. Specific antibody against LOL identified proteins of approximately 30, 42, 52 and 68 kd in various cells and in bovine aorta. These proteins were immunochemically distinct from the recombinant LO expressed by fibroblasts and from the bovine aorta LO. The LO gene (LOX) and LOXL were transiently up-regulated at early stages of liver granuloma development in Schistosoma mansoni-infected mice, although the peak of LOL mRNA synthesis preceded that of LO. LOL protein and LO were colocalized at sites of fibrogenesis in human lung fibrosis and in the stromal reaction of bronchiolo-alveolar carcinomas and of in situ ductal breast tumors. In conclusion, the LOL protein was identified as a secreted protein and localized in the extracellular matrix in active fibrotic diseases and in the early stromal reaction of breast cancer.

Topics: Adenocarcinoma, Bronchiolo-Alveolar; Animals; Carcinoma, Ductal, Breast; Cattle; Cell Line; Female; Fibrin; HeLa Cells; Humans; Liver Diseases, Parasitic; Lung; Lung Neoplasms; Mammary Neoplasms, Animal; Mice; Peptide Fragments; Protein-Lysine 6-Oxidase; Pulmonary Fibrosis; RNA, Messenger; Schistosomiasis mansoni; Stromal Cells

Topics: Animals; Anticoagulants; Blood Coagulation; Fibrin; Humans; Lung Neoplasms; Neoplasm Metastasis; Neoplasms; Platelet Aggregation Inhibitors

Topics: Animals; Aspirin; Blood Vessels; Capillaries; Endothelium; Fibrin; Heparin; Lung Neoplasms; Microcirculation; Neoplasm Metastasis; Neoplasms; Platelet Aggregation; Warfarin

Topics: Animals; Anticoagulants; Antigens, Neoplasm; BCG Vaccine; Blood Coagulation; Cell Adhesion; Cell Line; Cell Survival; Endothelium; Fibrin; Fibrinolysis; Humans; Immunity; Liver Neoplasms; Lung Neoplasms; Lymph Nodes; Macrophages; Mycobacterium bovis; Necrosis; Neoplasm Metastasis; Neoplasms; Organ Specificity; Platelet Aggregation

Topics: Blood Coagulation; Bone Marrow; Chronic Disease; Disseminated Intravascular Coagulation; Fibrin; Fibrinogen; Humans; Liver; Lung Neoplasms; Male; Prostatic Neoplasms; Surgical Procedures, Operative

Dense fibrin networks resistant to lysis have been reported in patients at high risk of thromboembolism. Little is known about fibrin clot properties in cancer. We investigated fibrin clot properties and their determinants in patients with inoperable lung cancer.. We enrolled 150 patients with advanced lung cancer prior to therapy and 90 control subjects matched by age, sex, cardiovascular disease, and diabetes. Plasma clot permeability (K. Advanced lung cancer is associated with the prothrombotic plasma clot phenotype largely driven by smoking.

Topics: Aged; Case-Control Studies; Cigarette Smoking; Cotinine; Female; Fibrin; Fibrin Clot Lysis Time; Humans; Lung Neoplasms; Male; Middle Aged; Phenotype; Thrombin

Lung cancer is the leading cause of cancer-related deaths worldwide, and lung squamous carcinomas (LUSC) represent about 30% of cases. Molecular aberrations in lung adenocarcinomas have allowed for effective targeted treatments, but corresponding therapeutic advances in LUSC have not materialized. However, immune checkpoint inhibitors in sub-populations of LUSC patients have led to exciting responses. Using computational analyses of The Cancer Genome Atlas, we identified a subset of LUSC tumors characterized by dense infiltration of inflammatory monocytes (IMs) and poor survival. With novel, immunocompetent metastasis models, we demonstrated that tumor cell derived CCL2-mediated recruitment of IMs is necessary and sufficient for LUSC metastasis. Pharmacologic inhibition of IM recruitment had substantial anti-metastatic effects. Notably, we show that IMs highly express Factor XIIIA, which promotes fibrin cross-linking to create a scaffold for LUSC cell invasion and metastases. Consistently, human LUSC samples containing extensive cross-linked fibrin in the microenvironment correlated with poor survival.

Topics: Animals; Biomarkers, Tumor; Carcinoma, Squamous Cell; Chemokine CCL2; Factor XIIIa; Female; Fibrin; Gene Expression Regulation, Neoplastic; Humans; Lung Neoplasms; Male; Mice; Mice, Inbred DBA; Monocytes; Neoplasm Invasiveness

Platelets are increasingly recognized for their contributions to tumor metastasis. Here, we show that the phosphoinositide signaling modulated by phosphatidylinositol transfer protein type α (PITPα), a protein which shuttles phosphatidylinositol between organelles, is essential for platelet-mediated tumor metastasis. PITPα-deficient platelets have reduced intracellular pools of phosphoinositides and an 80% reduction in IP

Topics: Animals; Anticoagulants; Blood Platelets; Fibrin; Gene Deletion; Hemostasis; Hyperplasia; Immunity, Mucosal; Inositol 1,4,5-Trisphosphate; Integrases; Lung Neoplasms; Lymphoid Tissue; Male; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Phospholipid Transfer Proteins; Platelet Aggregation; Signal Transduction; Thrombin; Thrombosis

Elevated interstitial fluid pressure and solid stress within tumors contribute to poor intratumoral distribution of nanomedicine. In this study, we hypothesized that the presence of fibrin in tumor extracellular matrix contributes to hindered intratumoral distribution of nanocarriers and that this can be overcome through the use of a fibrinolytic enzyme such as tissue plasminogen activator (tPA). Analysis of fibrin expression in human tumor biopsies showed significant fibrin staining in nearly all tumor types evaluated. However, staining was heterogeneous across and within tumor types. We determined the effect of fibrin on the diffusion, intratumoral distribution, and therapeutic efficacy of nanocarriers. Diffusivity of nanocarriers in fibrin matrices was limited and could be improved significantly by coincubation with tPA.

Topics: Albumins; Animals; Female; Fibrin; Fibrinolysis; Fibrinolytic Agents; Humans; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Nanomedicine; Paclitaxel; Tissue Plasminogen Activator; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Lung cancer is the most important causes of the cancer related mortality. Patients with lung cancer are usually diagnosed at advanced or locally advanced stage, for this reason early diagnosis of lung cancer is very important. For early detection of lung cancer some methods are emphasized such as low-dose computed tomography or tumor biomarkers. In this study we aimed to evaluate DR-70 sensitivity and specificity as a tumor marker in detection of non-small cell lung cancers.. Between May 2013 and April 2014, the serum samples from 88 non lung cancer patients, 86 patients with chronic obstructive pulmonary disesase were obtained. Blood samples from each participant were analyzed for DR-70 level.. Totally 174 patients were enrolled to the study (152 male, 22 female). Histopathologically 47(53.4%) patients were diagnosed with squamous cell lung cancer, 34 (38.6%) with adenocarcinoma, and 7 (8%) with non-small cell lung cancer. The mean serum DR-70 levels in lung cancer patients (2.43 ± 1.82 µg/mL) was significantly higher compared to the 86 non-cancerous subjects (1.15 ± 0.70 µg/mL) (p< 0.01). DR-70 exhibited clinical sensitivity and specificity of 54.5 and 83.7%, respectively, at an optimal cut off at 1.98 µg/mL. It could be said that the risk of the presence of the disease is 6.171 times higher in the cases where DR-70 level is 1.98 µg/mL and higher.. DR-70, a marker used to measure fibrin degradation products, generated by all major cancers, may helps to find high risk lung cancer patients.

Topics: Aged; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Humans; Immunoassay; Lung Neoplasms; Male; Middle Aged; Reproducibility of Results; ROC Curve; Tomography, X-Ray Computed

Previous studies have demonstrated that plasma D-dimer, a degradation product of cross-linked fibrin, correlated with tumor stage and prognosis in cancer patients. The aim of this study is to examine whether plasma D-dimer levels before and during chemotherapy predict tumor response and survival in advanced NSCLC patients undergoing first line chemotherapy.. Plasma D-dimer levels before (B0), and after one (B1) and two (B2) cycles of chemotherapy in 82 patients with advanced NSCLC were measured and correlated with treatment response, clinical features, progression-free survival (PFS) and overall survival.. A significant correlation was identified between changes in D-dimer levels before and after two chemotherapy cycles and treatment response. In addition, there were significant correlations between D-dimer positivity at B0, B1 and B2 time points and tumor stage, number of metastatic sites and treatment response. Patients with positivity of D-dimer at B0, B1 and B2 had significantly shorter PFS compared with those with negativity. Notably, positivity of D-dimer at B1 and B2 time points was an independent predictive factor for unfavorable PFS.. The positivity of D-dimer before and during chemotherapy is a predictor of treatment response and worse PFS in patients with advanced NSCLC. D-dimer levels provide prognostic information in addition to that of imaging studies.

Topics: Adult; Aged; Antineoplastic Agents; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cross-Linking Reagents; Disease-Free Survival; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Humans; Kaplan-Meier Estimate; Lung Neoplasms; Male; Middle Aged; Predictive Value of Tests; Prognosis; Treatment Outcome

Venous thromboembolism (VTE) is common in cancer patients, and is the second commonest cause of death associated with the disease. Patients with chronic inflammation, such as cancer, have been shown to have pathological clot structures with modulated mechanical properties. Fractal dimension (df) is a new technique which has been shown to act as a marker of the microstructure and mechanical properties of blood clots, and can be performed more readily than current methods such as scanning electron microscopy (SEM). We measured df in 87 consecutive patients with newly diagnosed lung cancer prior to treatment and 47 matched-controls. Mean group values were compared for all patients with lung cancer vs controls and for limited disease vs extensive disease. Results were compared with conventional markers of coagulation, fibrinolysis and SEM images. Significantly higher values of df were observed in lung cancer patients compared with controls and patients with extensive disease had higher values than those with limited disease (p< 0.05), whilst conventional markers failed to distinguish between these groups. The relationship between df of the incipient clot and mature clot microstructure was confirmed by SEM and computational modelling: higher df was associated with highly dense clots formed of smaller fibrin fibres in lung cancer patients compared to controls. This study demonstrates that df is a sensitive technique which quantifies the structure and mechanical properties of blood clots in patients with lung cancer. Our data suggests that df has the potential to identify patients with an abnormal clot microstructure and greatest VTE risk.

Topics: Aged; Algorithms; Biomarkers; Blood Coagulation; Blood Coagulation Tests; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Case-Control Studies; Female; Fibrin; Fractals; Hemorheology; Humans; Lung Neoplasms; Male; Microscopy, Electron, Scanning; Middle Aged; Neoplasm Staging; Prospective Studies; Risk; Single-Blind Method; Smoking; Thrombophilia; Venous Thromboembolism

Clusters of circulating tumor cells (CTC) exhibit more robust metastatic properties than single CTC. Thus, understanding the distinct behaviors of CTC clusters and how CTC clustering is regulated may offer new insights into how to limit metastasis. In this study, we utilized an in vivo confocal system to observe the clustering behavior of CTC in real time, finding that the number of clusters increased proportionally with the growth of the primary tumor. Our experiments also indicated that the flow rate of the CTC clusters in blood vessels was relatively slower than single CTC due to increased vessel wall adhesion. Depending on disease stage, 5% to 10% of total CTC in circulation were in clusters, with this proportion increasing to >24% within lung metastases examined. Notably, in the 4T1 mouse model of breast cancer metastasis, we found that injecting host animals with urokinase-type plasminogen activator, a clinical thrombolytic agent, was effective at preventing the assembly of CTC clusters and prolonging overall host survival by approximately 20% relative to control animals. Our results suggest a tractable approach to limit metastasis by suppressing the formation or stability of CTC clusters circulating in the blood of cancer patients.

Topics: Animals; Cell Adhesion; Cell Line, Tumor; Disease Models, Animal; Female; Fibrin; Fibrinolytic Agents; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Neoplasm Metastasis; Neoplastic Cells, Circulating; Urokinase-Type Plasminogen Activator

We recently introduced a technique of sutureless, mesh-based pneumostasis for preventing alveolar air leaks after lung resection. To verify the clinical usefulness of this technique, we examined if it can contribute to preserving gas exchange capacity and promoting postoperative rehabilitation.. We prospectively collected perioperative data, including arterial oxygen saturation on postoperative day (POD) 1 and the length of postoperative rehabilitation in 100 patients undergoing elective, video-assisted major lung resection for cancer. Before April, 2006, intraoperative air leaks were sealed with the conventional method (control group), and thereafter, with bioabsorbable mesh and glue, without suturing, (treated group). To reduce the bias in comparison of the nonrandomized control group, we paired the treated group with the control group using the nearest available matching method on the estimated propensity score.. Thirty-five patients in the control group were matched to 35 patients in the treated group based on the estimated propensity score. The length of both chest tube drainage and postoperative rehabilitation were significantly shorter in the treated group than in the control group (median, 1 versus 1 d, P = 0.03; 2 versus 3 d, P = 0.01, respectively). The arterial oxygen saturation on POD 1 was significantly higher in the treated group than in the control group (median, 94.0 versus 92.5 %, P = 0.03).. Mesh-based pneumostasis during video-assisted major lung resection enabled early chest tube removal, preserved postoperative oxygenation capacity, and promoted postoperative rehabilitation, which may facilitate fast-track surgery for patients undergoing video-assisted major lung resection for cancer.

Topics: Aged; Case-Control Studies; Chest Tubes; Device Removal; Exercise Tolerance; Female; Fibrin; Homeostasis; Humans; Lung; Lung Neoplasms; Male; Middle Aged; Pneumonectomy; Polyglycolic Acid; Prospective Studies; Pulmonary Gas Exchange; Retrospective Studies; Surgical Mesh; Treatment Outcome; Video-Assisted Surgery

The process of metastasis formation in cancer is not completely understood and is the main reason cancer therapies fail. Previously, we showed that dual liposomes simultaneously containing the hemostatic inhibitor, dipyridamole and the anticancer drug, perifosine potently inhibited metastasis, causing a 90% reduction in the number of lung metastases in a murine experimental metastasis model. To gain deeper insight into the mechanisms leading to the inhibition of metastasis by these dual liposomes, in the present study, the development of metastases by MT3 breast cancer cells in a mouse xenograft model was analyzed in more detail with regard to tumor cell settlement and metastatic growth. We found that the development of lung metastases by MT3 tumor cells is essentially dependent on the formation of fibrin clots as a precondition for the pulmonary arrest of tumor cells and the subsequent intravascular expansion of micrometastases before their invasion into the surrounding tissue.

Topics: Animals; Antineoplastic Agents; Blood Coagulation; Blood Vessels; Cell Line, Tumor; Dipyridamole; Female; Fibrin; Humans; Liposomes; Lung; Lung Neoplasms; Mammary Neoplasms, Experimental; Metallothionein 3; Mice; Mice, Nude; Neoplasm Metastasis; Phosphorylcholine; Platelet Aggregation Inhibitors; Xenograft Model Antitumor Assays

The attachment of circulating tumor cells to the blood vessels of distant organs is an important step in metastasis. We show here that experimental lung metastasis by two cell lines, B16F1 melanoma and 3LL lung carcinoma, is greatly reduced in transgenic mice that lack plasma fibronectin. This multifunctional adhesive glycoprotein becomes cross-linked to fibrin during clotting. Here, we report that eliminating plasma fibronectin from the blood circulation reverses the prometastatic effects of blood clotting and tumor cell integrin alphavbeta3. In vitro studies showed that fibrin-fibronectin complexes, but not purified fibrin, supported tumor cell attachment and invasion. These functions correlate with the ability of fibrin-fibronectin complexes to induce the activation of integrin alphavbeta3. Our findings reveal an important contribution of plasma fibronectin in lung metastasis. Furthermore, they suggest that the previously noted effects of blood clotting on lung metastasis might be mediated in part by a fibronectin-alphavbeta3 integrin axis, in which plasma fibronectin has to be incorporated into the blood clot.

Topics: Animals; Blood Coagulation; Carcinoma, Lewis Lung; Cell Adhesion; Fibrin; Fibronectins; Integrin alphaVbeta3; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neoplasm Invasiveness

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

A warfarin treated patient unexpectedly presented with an elevated international normalized ratio (INR). Repeat testing in two laboratories gave conflicting results. The chromogenic assay of factor X was used to determine the correct INR result. The patient had laboratory results consistent with a dysfibrinogenemia, which prevented detection of the endpoint with a photo-optical detection system. The chromogenic assay of factor X is recommended for monitoring patients on warfarin when the INR cannot be accurately determined due to interference with the fibrin endpoint in the INR.

Topics: Aged; Anticoagulants; Blood Coagulation; Chromogenic Compounds; Factor X; Female; Fibrin; Fibrinogen; Heart Failure; Humans; International Normalized Ratio; Lung Neoplasms; Prothrombin Time; Time Factors; Warfarin

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

Vascular endothelial growth factor (VEGF)-induced vascular permeability (VP) is a hallmark of tumor growth and metastasis. Previous studies have shown a requirement for Src kinase in VEGF-mediated VP and signaling in blood vessels. In this study, we have examined the effect of Src-mediated reduced VP on tumor growth and metastasis. The growth and spontaneous metastasis of VEGF-expressing tumor cells were determined in Src-knockout (src(-/-)) or control mice (src(+/+) or src(+/-)). In comparison to control mice, src-null mice had a significant reduction in tumor-induced VP as well as a subsequent reduction in spontaneous metastasis. In contrast, primary tumor weight and vascular density were unchanged between src-null and control mice. Consistent with a role for Src in the extravasation of tumor cells from the circulation, direct intravenous injection of lung carcinoma cells resulted in a more than 2-fold reduction in lung tumor burden in src-null mice compared to control mice. The comparison of the results from the experimental metastasis and the spontaneous metastasis models suggests that there are defects in VP in the primary site of Src-deficient mice and that there may be an essential role for Src and Src-mediated VP in tumor metastasis to the lung.

Topics: Animals; Capillary Permeability; Carcinoma, Lewis Lung; Cell Line, Tumor; Cell Proliferation; Fibrin; Fibrin Fibrinogen Degradation Products; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Mice, Knockout; Neoplasm Metastasis; Neovascularization, Pathologic; src-Family Kinases; Vascular Endothelial Growth Factor A

Comprehensive studies of fibrinolysis in non-small cell lung carcinoma have been limited, and assignment of patients to high/low prognosis groups based on arbitrary cut-offs utilizing fibrinolytic measurements is unstandardized. This study was performed in 166 patients to examine the effects of cut-off values determined in three ways. Model 1 assigned patients to one of three equal groups (low, medium, high) based on fibrinolytic measurements made at diagnosis, Model 2 divided patients into low/high groups using median values, and Model 3 grouped according to the parameter being above/below normal range. In model 1, raised plasma fibrinogen, D-dimer and soluble fibrin were positively associated with poorer survival. In model 2, tissue plasminogen activator antigen was additionally related to poorer prognosis. Model 3 identified seven parameters as significantly related to survival, two not identified by the other models becoming significant [plasmin-antiplasmin, tissue plasminogen activator inhibitor-1 (PAI-1) antigen]. Using multivariate analysis, plasma fibrinogen level was the most uniformly significant parameter. Relative risk estimates indicated that raised plasma fibrinogen, soluble fibrin and D-dimer were associated with increased risk of death. Use of the normal/above normal cut-off is recommended to provide the maximum number of significant parameters relating to prognosis, and increased plasma D-dimer, PAI-1 antigen and fibrinogen were most closely related to survival/prognosis.

Topics: Adult; Aged; Aged, 80 and over; Antifibrinolytic Agents; Biomarkers; Carcinoma, Non-Small-Cell Lung; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Fibrinogen; Fibrinolysis; Humans; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Plasminogen Activator Inhibitor 1; Prognosis; Reference Values; Risk Factors; Serine Proteinase Inhibitors; Survival Rate

There is considerable evidence for a relationship between hemostasis and malignancy. Since platelet adhesion to tumor cells has been implicated in the metastatic process and plasma levels of fibrinogen (Fg) and soluble fibrin (sFn) monomer are increased in cancer, we hypothesized that these molecules might enhance tumor-platelet interaction. We therefore studied binding of sFn monomer to tumor cells in a static microplate adhesion assay and determined the effect of pre-treating tumor cells with sFn on tumor cell-induced thrombocytopenia and experimental metastasis. Soluble fibrin (produced by adding thrombin to FXIII- and plasminogen-free Fg in the presence of Gly-Pro-Arg-Pro-amide (GPRP-NH2) significantly increased platelet adherence to tumor cells. This effect was primarily mediated by the integrins alphaIIb beta3 on the platelet and CD 54 (ICAM-1) on the tumor cells. Platelets adhered to untreated A375 cells (28 +/- 8 platelets/tumor cell) and this was not significantly affected by pre-treatment of the tumor cells with fibrinogen or GPRP-NH2. Although thrombin treatment increased adherence, pre-incubation of the tumor cells with sFn resulted in a further increase in platelet binding to tumor cells. In contrast to untreated tumor cells, intravenous injection of sFn-treated A 375 cells reduced the platelet count in anticoagulated mice, supporting the in vitro finding that sFn enhanced tumor cell-platelet adherence. In a more aggressive model of experimental metastasis, treating tumor cells with sFn enhanced lung seeding by 65% compared to untreated cells. Extrapolation of our data to the clinical situation suggests that coagulation activation, and subsequent increase in circulating Fn monomer, may enhance platelet adhesion to circulating tumor cells and thereby facilitate metastatic spread.

Topics: Animals; Antigens, CD; Antigens, Human Platelet; Batroxobin; Blood Platelets; Cell Adhesion; Cell Communication; Female; Fibrin; Fibrinolytic Agents; Flow Cytometry; Hemostatics; Humans; Lung Neoplasms; Melanoma, Amelanotic; Mice; Mice, Nude; Platelet Adhesiveness; Receptors, Thrombin; Solubility; Thrombin; Thrombocytopenia; Tumor Cells, Cultured

Involvements of interleukin-6 (IL-6) and fibrinogen in cancer development have been independently studied. However, the association of these molecules in cancer patients remains uncertain. This study was conducted to clarify the association according to the clinicopathological characteristics of lung cancer patients.. Serum IL-6 levels assayed in 339 patients without pleural effusion were assessed according to clinical stage, histological type of the cancer and levels of fibrin (ogen) degradation products (FDP), and C-reactive protein (CRP).. Elevations of serum IL-6 levels more than 4 pg/ml were found in 37.8% of all patients. According to the clinical stage and histological type, the elevations were significantly more frequent in the advanced stage (44.7%), in squamous cell (49.1%) and large cell carcinomas (63.6%). Similarly, the frequency of the elevated cases (> 400 mg/dl) and the mean value of the fibrinogen level were also higher in the advanced stage (54.2%, 455.0 mg/dl) and large cell carcinoma (54.6%, 459.3 mg/dl), respectively. The elevations of fibrinogen, FDP and CRP levels were found to be related to those of the IL-6 level.. In lung cancer, serum IL-6 elevations are particularly frequent in the advanced stages of patients with squamous cell and large cell carcinoma, which are associated with the elevated levels of fibrinogen, suggesting a possibility that IL-6 was involved not only directly, but also indirectly, through regulating plasma fibrinogen with promotion of cancer development in vivo.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; C-Reactive Protein; Carcinoma, Large Cell; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Fibrinogen; Humans; Interleukin-6; Lung Neoplasms; Male; Middle Aged

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

Extravascular, intratumoral fibrin deposition is frequently observed within and around neoplastic tissue and has been implicated in various aspects of tumour growth. This is the first report on the presence and distribution of fibrinogen/fibrin in primary (14 glioblastomas) and metastatic (nine samples of lung cancer origin) human brain tumours detected by immunofluorescent techniques. All tissue samples showed specific staining for fibrinogen/fibrin. In glioblastomas fibrin deposits could be detected within and around tumour foci, while in metastatic brain tumours the tumour cell nodules were surrounded by fibrin deposits localized almost exclusively in the connective tissue compartment of tumours. Double-labelling reactions for von Willebrand factor and fibrinogen/fibrin has revealed that fibrin deposition occurred throughout the tumour stroma independently of tumour vasculature. The overlapping reactions for fibrinogen/fibrin and factor XIII subunit A, as well as the urea-insolubility of the deposits indicate the crosslinked, highly stabilized nature of fibrin both within and around tumours. Staining with Ki M7 monoclonal antibody specific for phagocytosing macrophages showed these cells to be scattered in the nonnecrotic areas in glioblastomas and to be accumulated at the interface of tumorous parenchyma and connective tissue in both primary and metastatic tumours. The close association between fibrin deposition and macrophage accumulation strongly suggests the active participation of tumour associated macrophages in the formation of stabilized intratumoral fibrin network in human brain neoplasms.

Topics: Blood Coagulation; Brain Neoplasms; Connective Tissue; Factor XIII; Fibrin; Fibrinolysis; Glioblastoma; Humans; Lung Neoplasms; Macrophages; Neoplasm Proteins

Tumor cells interact with the hemostatic system in various ways and may thus influence malignant growth and spread. MC28 fibrosarcoma cells possess a potent procoagulant activity (PCA) and form lung tumors following intravenous injection. The aim of this work was to study the relationship between PCA, intravascular coagulation and lung seeding in the MC28 model. MC28 cells were injected into control, warfarinized and heparinized hooded Lister rats. Coagulation changes were monitored by thromboelastography (TEG) and Sonoclot analysis (SA), lung fibrin formation by light and electron microscopy, tumor seeding by macroscopic counting and tumor cell and platelet deposition in the lungs by radiolabelling. PCA was measured by chromogenic assay. MC28 PCA was characterized as a tissue factor-factor VIIa complex that probably arose during cell culture or disaggregation of solid tumors. Injection of tumor cells caused marked coagulopathy and was rapidly (within 30 min) followed by fibrin deposition in the lungs and accumulation of radiolabelled platelets. Heparin and warfarin significantly reduced lung seeding (p < 0.001) and reduced retention of radiolabelled tumor cells in the pulmonary circulation (p < 0.01). Inhibition of cellular PCA by prior treatment with concanavalin A markedly reduced intravascular coagulation and lung seeding. We conclude that MC28 cells cause intravascular coagulation as a direct result of their procoagulant activity. The data suggest that tumor cells form complexes with platelets and fibrin which are retained in the lungs long enough for extravasation and seeding to occur.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Blood Coagulation; Blood Coagulation Tests; Blood Platelets; Cell Adhesion; Concanavalin A; Culture Media, Serum-Free; Cysteine Endopeptidases; Disseminated Intravascular Coagulation; Fibrin; Fibrosarcoma; Heparin; Injections, Intravenous; Lung; Lung Neoplasms; Macrophage Activation; Male; Neoplasm Proteins; Neoplasm Transplantation; Neoplastic Cells, Circulating; Pulmonary Circulation; Rats; T-Lymphocytes; Tumor Cells, Cultured; Warfarin

Intrapleural fibrin deposition commonly accompanies pleural injury and may contribute to the organization of exudative pleural effusions, which leads to lung entrapment. Previous investigators have observed an increase in procoagulant proteins in pleural effusions but very little thrombin formation. FVa is the protein cofactor in the prothrombinase complex that dramatically enhances the generation of thrombin from prothrombin by the serine protease fXa. The presence of fVa within the pleural space could influence fibrin formation and pleural scarification. We examined pleural fluids obtained from patients who had lung cancer, CHF, and empyema for the presence of fV/fVa. The fV antigen was increased in exudative pleural fluids, in comparison with transudates. However, the specific activity of fV antigen present in exudates was significantly less than that observed for the lower concentration of antigen present in transudate and could not be activated to the same degree by thrombin. Immunoblots of fV antigen in exudates indicated that fV was partially cleaved and inactivated by unidentified proteases. We conclude that although fV is present in pleural fluid, it may be present in a degraded form, which may partially account for a lack of thrombin-generating capacity in these pleural fluids. The presence of fV does not necessarily correlate with pleural loculation.

Topics: Blood Coagulation Factors; Empyema; Factor V; Fibrin; Fibrinolysin; Glucose; Heart Failure; Humans; Immunoblotting; L-Lactate Dehydrogenase; Lung Neoplasms; Molecular Weight; Pleural Effusion; Thrombin

We examined the effect of fibrin coagulation on tumor cytotoxicity mediated by human lymphokine (IL-2)-activated killer (LAK) cells. LAK cells were induced from peripheral blood mononuclear cells (MNC) by culture with recombinant IL-2 for 4 or 5 days, and LAK cell-mediated cytotoxicity against tumor cells was assessed by 51Cr release assay in the presence or absence of plasma from normal subjects and lung cancer patients. Plasma did not affect the phase of induction of LAK activity by IL-2, but dose-dependently inhibited the effector phase of LAK cell-mediated cytotoxicity against Daudi cells. Similar inhibition of LAK cell-mediated cytotoxicity was observed on pretreatment of Daudi cells and human lung cancer cell lines with human fibrinogen plus thrombin. A parallel relationship was found between the amount of fibrinogen in plasma of lung cancer patients and inhibition of LAK cytotoxicity. This inhibition was reduced by addition of anticoagulants (heparin or argatroban). These findings suggest that fibrin coagulation on tumor cells protects them from LAK cell-mediated tumor cytotoxicity in malignant lesions and that a combination of an anticoagulant drug and IL-2/LAK therapy may be effective for treatment of lung cancer patients.

Topics: Aged; Aged, 80 and over; Antithrombins; Arginine; Blood Coagulation; Cell Death; Cytotoxicity, Immunologic; Female; Fibrin; Fibrinogen; Humans; Immunity, Cellular; Interleukin-2; Killer Cells, Lymphokine-Activated; Kinetics; Leukocytes, Mononuclear; Lung Neoplasms; Male; Middle Aged; Pipecolic Acids; Sulfonamides; Thrombin

Extravascular coagulation and fibrinolysis are intimately involved in and modulate cancer cell growth, invasion and metastasis. Samples from resection specimens of patients with primary lung cancer (adenocarcinomas) were tested with monoclonal (MAb) and polyclonal (PAb) antibodies against various factors of the coagulation or fibrinolysis systems, or against antigens of inflammatory or proliferating cells. MAb Ki-67 specific to nuclear antigens of proliferating cells showed a distinct but variable staining of cell nuclei throughout the tumor tissue. Nests of tumor tissue stained with cytokeratin-specific antibodies (PKK1), whereas other parts were negative. Fibrin(ogen) and fibronectin were found throughout the tumor tissue stroma and in the alveolar lining, and the most densely stained areas were at the transition zone between normal and tumor tissue. Fibrinolytic system components like tissue plasminogen activators (t-PA), and urokinase (u-PA), and their inhibitors PAI-1 and PAI-2 were all studied. All specimens were negative for t-PA (except endothelial linings), whereas urokinase-specific antibodies stained loosely packed tumor cells and macrophages within the tumor stromal tissue and alveolar septa. Both PAI-1 and PAI-2 were most prominently expressed within interstitial and alveolar macrophages. A weaker staining of tumor tissue cells was demonstrated. Inflammatory cells like macrophages and T lymphocytes were located in aggregates or diffusely spread within tumor stromal tissue. The inflammatory reaction was most intense at the border between normal lung and tumor tissue.

Topics: Adenocarcinoma; Antibodies; Antibodies, Monoclonal; Biomarkers; Blood Coagulation; Fibrin; Fibrinogen; Fibrinolysis; Fibronectins; Humans; Immunohistochemistry; Inflammation; Keratins; Ki-67 Antigen; Lung Neoplasms; Macrophages; Nuclear Proteins; Plasminogen Inactivators; T-Lymphocytes; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

A relationship between blood coagulation system and tumor growth is discussed. The role of hemocoagulation disorders in the pathogenesis, dissemination and advancement of disease at molecular, cellular, body and population level is discussed on the basis of the authors' results and literature data. Rastr electron microscopy and immunofluorescent method revealed fibrin on the surface of tumor cells both in vivo and in vitro which is thought to play a masking and protective role. Studies of clotting and anticlotting factors in various subgroups of healthy subjects and patients with precancer and cancer revealed a steady-state increase in blood coagulability in cancer patients of old age. On these grounds, blood fibrinogen level was used for screening. Increased fibrinogen level was associated with higher tumor occurrence. The authors' concept of the pathogenetic role of the coagulation system in tumor growth provided a rationale for the use of direct and indirect anticoagulants in addition to cytotoxic drugs for breast, ovarian and gastric cancer. As a result, cytotoxic treatment was more effective and better tolerated.

Topics: Adult; Anticoagulants; Antineoplastic Agents; Blood Coagulation Disorders; Breast Neoplasms; Cell Line; Cells, Cultured; Drug Therapy, Combination; Female; Fibrin; Fibrinogen; Humans; Lung Neoplasms; Male; Middle Aged; Stomach Neoplasms; Surface Properties; Tumor Cells, Cultured

Mechanisms of coagulation activation in situ were studied by means of immunohistochemical techniques applied to surgically resected primary adenocarcinomas and squamous cell carcinomas of the lung. Findings in these two histologic types were similar. Double-labeling techniques using macrophage-specific antibody together with antibody to either tissue factor, factor VII, factor X, or factor V revealed coincident staining for each of these coagulation factors on tumor-associated macrophages. Staining of tumor cells for these factors was rare and inconsistent. Both macrophages and fibroblasts in the tumor connective tissue stained for the a subunit of factor XIII. Fibrinogen was abundant throughout the tumor connective tissue, but staining for fibrin and D-dimer cross-linked sites of fibrin was restricted to areas adjacent to macrophages, indicating that thrombin was generated in association with tumor macrophages but not with tumor cells. By contrast, tumor cells stained diffusely for urokinase-type plasminogen activator and focally for thrombomodulin. These findings contrast with those reported previously for small cell carcinoma of the lung and suggest that coagulation activation in adenocarcinoma and squamous cell carcinoma of the lung may occur indirectly through activation of certain host cells such as macrophages. By contrast, tumor cell plasminogen activator may mediate certain aspects of the malignant phenotype in these tumor types.

Topics: Adenocarcinoma; Antibody Specificity; Antigens, Neoplasm; Blood Coagulation; Carcinoma, Squamous Cell; Fibrin; Fibrinolysis; Humans; Immunohistochemistry; Lung Neoplasms; Macrophages; Thrombin; Urokinase-Type Plasminogen Activator

Components of coagulation and fibrinolysis reactions were identified in situ by immunohistochemical staining in fresh frozen sections of small cell carcinoma of the lung tissue. Tumor cells stained positively for tissue factor, a protein that is capable of activating the extrinsic pathway of coagulation (the components of which have been seen within small cell carcinoma of the lung [SCCL] tissue), and for proteins C and S antigens. Fibrin was seen in a focal distribution at the host-tumor interface, indicating that thrombin had acted upon the fibrinogen found throughout the tumor stroma. Staining with a neoepitope-specific antibody, which does not discriminate between fibrinogen fragment D and fibrin fragment D-dimer, was similar to that of the fibrin antibody. High molecular weight urokinase-type and tissue-type plasminogen activators were seen in vascular endothelium, but neither existed within the tumor. Low molecular weight urokinase was found in rare isolated foci of tumor cells primarily adjacent to areas of necrosis. Plasminogen activator inhibitor-3 occurred in tumor cell cytoplasmic blebs and in necrotic tumor cells, but plasminogen activator inhibitors 1 and 2 were not seen. Our data suggest a mechanism for thrombin generation and fibrin formation within SCCL tissues that could support cell proliferation, stroma formation, and preservation. These features could be conductive to perpetuation of this tumor and conceivably could form the basis of the beneficial effects of antithrombotic therapy seen in SCCL.

Topics: Blood Coagulation; Carcinoma, Small Cell; Factor V; Fibrin; Fibrinogen; Fibrinolysis; Glycoproteins; Humans; Immunohistochemistry; Lung Neoplasms; Plasminogen Activators; Plasminogen Inactivators; Protein C; Protein S

Erythrocyte CR1, a C3b/C4b-binding complement-regulatory protein, is sensitive to proteolysis in vitro. To test the hypothesis that in vivo erythrocyte CR1 reduction results from intravascular proteinase activities, we used enzyme-linked immunosorbent assays to measure gamma-crosslinked fibrin degradation products (D-dimers) as indicators of coagulation/fibrinolytic activity, and complexes of neutrophil elastase with alpha 1 proteinase inhibitor (E/A) as indicators of neutrophil enzyme release in malignant and inflammatory disorders. Erythrocyte CR1, measured by monoclonal anti-CR1 antibody binding, was inversely related to disease activity and blood proteinase markers. Levels of erythrocyte CR1 were significantly lower for patients with active versus remittent squamous and small cell lung cancers, Hodgkin's and diffuse large cell lymphomas, and acute myelogenous leukemias. In patients with active thoracic cancers, elevated D-dimer levels correlated with reduction of CR1. In patients with rheumatoid arthritis, CR1 reduction was correlated with elevated levels of elastase complexes. Our findings substantiate the relationship of acquired CR1 reduction to the activity of certain diseases and provide circumstantial support for the hypothesis that erythrocyte CR1 is lost to proteolysis in vivo. Although heritable differences in CR1 expression reduce the interpretability of single measurements of erythrocyte CR1 levels, disease-associated CR1 reduction may be a useful indicator of disorders with chronically increased blood proteinase activity.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Carcinoma, Small Cell; Erythrocyte Membrane; Fibrin; Humans; Lung Neoplasms; Middle Aged; Peptide Hydrolases; Receptors, Complement

In order to characterize further the previously observed induction of a highly metastatic phenotype in mouse bladder carcinoma cells by Ha-ras transfection, we studied production of plasminogen activator, in vitro invasiveness, and the potential for lung colonization of these cells. The parent carcinoma cells produced predominantly tissue-type plasminogen activator. Out of 13 clones of ras-transfected cells tested, 8 secreted quantitatively elevated levels of plasminogen activator (up to 3.5-fold) as compared to the control transfectants. The plasminogen activator activity in cell lysates was maximally increased 3-fold, the surface-associated activity increased 2.5-fold. The secreted plasminogen activator of cloned ras-transfected cells was characterized to be predominantly of the urokinase type (71.3% compared to 20.5% with the parental BL cells). Thus, in addition to the quantitative augmentation of plasminogen activator production and secretion in a large fraction of the ras-transfected cell population, a significant qualitative shift from tissue-type to urokinase-type has been observed. In addition, ras-transfection augmented the capacity of the cells for invasion into Matrigel in a double-filter in vitro assay as well as their ability to colonize the lungs of syngeneic animals. These malignant properties of the transfected cells might be responsible for their highly metastatic behaviour induced by ras transfection.

Topics: Animals; Fibrin; Genes, ras; Humans; Lung Neoplasms; Mice; Neoplasm Invasiveness; Neoplasm Metastasis; Phenotype; Transfection; Tumor Cells, Cultured; Urinary Bladder Neoplasms; Urokinase-Type Plasminogen Activator

Thrombin-generated cleavage sites of human fibrinogen have been identified adjacent to viable tumor cells in fresh frozen sections of small cell carcinoma of the lung (SCCL) by means of immunohistochemical techniques using mouse monoclonal antibodies to the N-terminal peptides of the fibrinogen alpha and beta chains. These results indicate that thrombin is generated in situ in this tumor type. Based on previous evidence for the existence of an initiator of coagulation together with coagulation factor intermediates associated with viable SCCL tumor cells in situ, and also for the favorable effects of anticoagulant therapy with warfarin in SCCL, we postulate that local tumor cell-induced thrombin formation may contribute to self-regulated progression of SCCL through deposition of fibrin and stimulation of cell proliferation. These results suggest novel treatment strategies for this particular tumor type and justify efforts to identify other tumor types in which similar mechanisms exist.

Topics: Antibodies, Monoclonal; Carcinoma, Small Cell; Fibrin; Fibrinogen; Humans; Immunoenzyme Techniques; Lung Neoplasms

Coumarins inhibit metastasis in a number of animal models, but the mechanism of this effect remains unclear. We have investigated the relationship between the coagulation system and metastasis using a new model system, involving i.v. injection of Mtln3 rat mammary carcinoma cells into Fischer 344 rats, and subsequent estimation of pulmonary seeding. Injection of factors II, VII, IX and X elevated the median number of surface pulmonary seedlings per animal to 182, and injection of factors II, IX and X to 181, compared with a median for control animals of 12 (P less than 0.001). Injection of factor VII alone, or of bovine serum albumin did not significantly affect pulmonary seeding. In a second experiment, arvin defibrination reduced the mean plasma fibrinogen concentration to 76.8 mg dl-1 from a control value of 228 mg dl-1. This degree of defibrination had no significant effects on pulmonary seeding, nor on the enhancing effects of factor complex injection (median numbers of seedlings per animal; control 15, arvin 21, arvin plus factors II, VII, IX and X 170, factors II, VII, IX and X only, 157). Factor complex injections did not detectably shorten thrombotest clotting times. In vitro testing suggested that Mtln3 cells contain little or no conventional factor X activating cancer procoagulant. The complex of coagulation factors II, IX and X appears to contain a component which greatly enhances metastasis in this model. This may explain the previously reported antimetastatic effect of coumarin anticoagulants, which suppress factors II, VII, IX and X. The enhancing effect of the factor complex does not appear to be altered by significant reductions in fibrin forming capacity, and defibrination itself has no effect on metastasis. These findings suggest the possibility that the effect of this factor complex on metastasis may be mediated via mechanisms other than the formation of a fibrin clot.

Topics: Ancrod; Animals; Anticoagulants; Blood Coagulation Factors; Female; Fibrin; Fibrinogen; Lung Neoplasms; Mammary Neoplasms, Experimental; Neoplasm Metastasis; Rats; Rats, Inbred F344; Tumor Cells, Cultured

Four cases of lung cancer were analysed for aspects relating to optical light microscopy and ultrastructure of coagulation disorders. Criteria of intravasal coagulopathy were positively verified from all four cases and were found to have grown manifest in the form of prethrombi in veins, arterioles, and venules, masses of fibrin monomer in capillaries, and the sludge phenomenon. Extravasation of fibrin was also observed around highly differentiated cancer cells as well as in tumour stroma and in necrotic foci. Most of the fibrin was identified in the forms of monomers and their ultrastructural variants. Morphological criteria of locally delimited coagulation disorders in the microcirculation of tumours are discussed together with the role played by fibrin in growth and dissemination of neoplasms.

Topics: Arteries; Blood Coagulation Disorders; Fibrin; Humans; Lung Neoplasms; Male; Microcirculation; Middle Aged; Thrombosis; Veins

Human recombinant single chain urokinase-type plasminogen activator (recombinant scu-PA) and a hybrid between human tissue-type plasminogen activator (t-PA) and scu-PA, obtained by ligation of cDNA fragments encoding the NH2-terminal region (amino acids 1-67) of t-PA and the COOH-terminal region (amino acids 136-411) of scu-PA, were expressed in a mammalian cell system. The proteins were purified from conditioned culture media containing 2% fetal calf serum by chromatography on zinc chelate-Sepharose, immunoadsorption chromatography on an insolubilized murine monoclonal antibody directed against urokinase, benzamidine-Sepharose chromatography, and Ultrogel AcA 44 gel filtration. Between 180 and 230 micrograms of the purified proteins were obtained per liter of conditioned medium, with a yield of approximately 18% and a purification factor of 720-1900. On sodium dodecyl sulfate gel electrophoresis under reducing conditions, the proteins migrated as single bands with approximate Mr 50,000 for recombinant scu-PA and Mr 43,000 for the t-PA/scu-PA hybrid. Following conversion to urokinase with plasmin, the proteins had a specific amidolytic activity comparable to that of natural scu-PA. Both proteins activated plasminogen directly with Km = 0.53 and 1.4 microM and k2 = 0.0034 and 0.0027 s-1, respectively. Both proteins did not bind specifically to fibrin and had a comparable degree of fibrin selectivity as measured in a system composed of a whole human 125I-fibrin-labeled plasma clot suspended in human plasma. It is concluded that this chimeric protein, consisting of the NH2-terminal "finger-like" domain of t-PA and the COOH-terminal region of scu-PA, has very similar enzymatic properties as compared to scu-PA, but has not acquired the fibrin affinity of t-PA.

Topics: Adenocarcinoma; Amino Acid Sequence; Amino Acids; Animals; Antibodies, Monoclonal; Chemical Phenomena; Chemistry, Physical; Cricetinae; DNA; Fibrin; Humans; Kinetics; Lung Neoplasms; Molecular Weight; Recombinant Fusion Proteins; Recombinant Proteins; Structure-Activity Relationship; Substrate Specificity; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

Topics: Animals; Dogs; Drug Combinations; Embolization, Therapeutic; Fibrin; Iodized Oil; Lung Neoplasms; Pulmonary Artery

A potential synergic effect of tissue-type plasminogen activator (t-PA), single-chain urokinase-type plasminogen activator (scu-PA) or urokinase on clot lysis was investigated in a whole human plasma system in vitro. The system consisted of a human plasma clot labeled with 125I-fibrinogen, immersed in citrated whole human plasma, to which the thrombolytic agents were added. Clot lysis was quantitated by measurement of released 125I, and activation of the fibrinolytic system in the surrounding plasma by measurements of fibrinogen and alpha 2-antiplasmin. t-PA, scu-PA and urokinase induced a dose-dependent and time-dependent clot lysis; 50 percent lysis after 2 h was obtained with 5 nM t-PA, 20 nM scu-PA and 12 nM urokinase. At these concentrations no significant activation of the fibrinolytic system in the plasma was observed with t-PA and scu-PA, whereas urokinase caused significant alpha 2-antiplasmin consumption and concomitant fibrinogen degradation. The shape of the dose-response curves was different; t-PA and urokinase showed a log linear dose-response whereas that of scu-PA was sigmoidal. Combinations of t-PA and scu-PA, of t-PA and urokinase or of scu-PA and urokinase at thrombolytic doses of each showed no synergism for thrombolysis. Fifty percent clot lysis in 2 h was obtained at total concentrations of the combined agents of 5 to 15 nM with molar ratios ranging from 1:4 to 4:1.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Drug Synergism; Fibrin; Fibrinolysis; Humans; Kinetics; Lung Neoplasms; Melanoma; Plasminogen Activators; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

A low Mr form (Mr 32,000) of single-chain urokinase-type plasminogen activator (scu-PA) was isolated from conditioned culture medium of a human lung adenocarcinoma cell line, CALU-3 (ATCC, HTB-55). The purified material (scu-PA-32k) consists of a single polypeptide chain and is immunologically similar to Mr 33,000 urokinase. Its NH2-terminal sequence is identical to that beginning at Leu-144 of Mr 54,000 urokinase. Whereas low Mr urokinase is derived from mature Mr 54,000 scu-PA by limited hydrolysis by plasmin first of the Lys-158-Ile-159 peptide bond and then of the Lys-136-Lys-137, scu-PA-32k is generated by specific hydrolysis of the Glu-143-Leu-144 peptide bond by an unidentified protease. scu-PA-32k resembles its Mr 54,000 scu-PA counterpart by its very low activity on chromogenic substrates for urokinase, by plasminogen-dependent fibrinolytic activity on fibrin plates, and by the lack of specific binding to fibrin. It activates plasminogen directly with high affinity, Km = 0.9 microM, but low turnover number, kcat = 0.0028 s-1. It is converted to fully active two-chain urokinase by plasmin with Km = 12 microM and kcat = 0.3 s-1. Like Mr 54,000 scu-PA, it causes significant lysis of a 125I-labeled fibrin clot in human plasma with relatively less fibrinogen breakdown as compared to urokinase. scu-PA-32k, which also has conserved fibrin specificity, represents a molecular variant which may be more suitable for large scale production as a fibrin-specific thrombolytic agent by recombinant DNA technology.

Topics: Adenocarcinoma; Amino Acid Sequence; Cell Line; Fibrin; Fibrinogen; Humans; Immunodiffusion; Kinetics; Lung Neoplasms; Molecular Weight; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

Acquired dysfibrinogenemia has not been previously reported as a paraneoplastic marker for malignancy. This report describes the clinical course of a patient who at the time of diagnosis of nonmetastatic renal cell carcinoma had dysfibrinogenemia characterized by prolongation of the thrombin and Reptilase times and increased sialic acid content of the purified fibrinogen. The thrombin and Reptilase times returned toward normal values after nephrectomy but became abnormal with the development of nonhepatic metastases. It is concluded that acquired dysfibrinogenemia can be part of a paraneoplastic syndrome and is a sensitive plasma marker for tumor progression.

Topics: Carcinoma, Renal Cell; Female; Fibrin; Fibrinogen; Follow-Up Studies; Humans; Kidney Neoplasms; Lung Neoplasms; Middle Aged; N-Acetylneuraminic Acid; Nephrectomy; Paraneoplastic Syndromes; Sialic Acids; Thrombin Time

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

Lewis lung carcinoma cells were implanted in the foot-pads of mice and the effects of the plasminogen-plasmin inhibitor tranexamic acid (t-AMCHA) and of the plasminogen activator urokinase on metastasis were examined by electron microscopy. The intravascular tumour cells were not associated with thrombus formation in either control or urokinase-treated mice. Polymerized fibrin deposition around tumour cells and thrombi composed of fibrin and platelets was observed only in the mice given t-AMCHA. This suggests that the inhibition of fibrinolysis by tACC caused fibrin deposition and thrombus formation around intravascular tumour cells, which prevented release of the cells from primary foci to form secondary tumours. On the other hand, fibrinolysis induced by urokinase prevented thrombus formation, and accelerated cell release from primary foci.

Topics: Animals; Blood Platelets; Cyclohexanecarboxylic Acids; Endopeptidases; Fibrin; Lung Neoplasms; Male; Mice; Mice, Inbred Strains; Microscopy, Electron; Neoplasm Transplantation; Neoplasms, Experimental; Thrombosis; Tranexamic Acid; Urokinase-Type Plasminogen Activator

This study was made to know the significance of fibrinopeptide A(FPA) as an indicator for coagulative analysis in thrombotic diseases. In normal control subjects (n=21), values of FPA by the radioimmunoassay were 0.5 +/- 1.4 ng/ml (mean +/- SD). In animal models, using Lyoplastin (tissue thromboplastin, n=5) or Ancrod (n=5) to piglets, plasma FPA levels elevated rapidly as a reflection of fibrin formation, and these changes of FPA were found to be most rapid and sensitive among the indicators for coagulation and fibrinolysis. In patients with thrombosis (n=32), elevated FPA levels (14.7 +/- 13.8 ng/ml) and beta-thromboglobulin (beta-TG)(86.1 +/- 65.6 ng/ml) were found. FPA levels in these patients positively correlated to beta-TG (r=0.5539, P less than 0.05) and inversely to fibrinogen (fbg) (r= -0.3622, P less than 0.05). In patients with acute myelocytic leukemia (AML, n=112), acute promyelocytic leukemia (APL, n=18) and acute lymphocytic leukemia (ALL, n=15), mean FPA levels in patients with active signs and symptoms were significantly higher (AML: 13.5 ng/ml, APL: 20.8 ng/ml, ALL: 12.4 ng/ml) than those examined during remission states (AML: 7.7 ng/ml, P less than 0.02, APL: 3.9 ng/ml, P less than 0.01, ALL: 2.7 ng/ml, P less than 0.01). FPA levels in patients with APL inversely correlated to fbg (r= -0.6399, P less than 0.01). In patients with lung cancer (n=75), mean FPA level in advanced stage (17.7 ng/ml, n=67) were significantly higher than those examined in early stage 6.5 ng/ml, n=8, P less than 0.001). In patients with acute disseminated intravascular coagulation (n=12), prolonged prothrombin time and activated partial thromboplastin time, severely reduced fbg and platelets, and remarkably elevated fibrin degradation product were found. Elevated FPA and beta-TG levels were also found (FPA: 23.5 +/- 15.0 ng/ml, beta-TG: 100.0 +/- 63.0 ng/ml). In five patients with thrombotic diseases who were treated successfully with 12500 IU of heparin per 12 hours (subcutaneous injection), plasma FPA levels were reduced to near normal levels quicker than changes of other indicators. These clinical and experimental data suggested that FPA was an useful indicator for active coagulation process.

Topics: Adult; Animals; Blood Coagulation; Disseminated Intravascular Coagulation; Female; Fibrin; Fibrinogen; Fibrinopeptide A; Heparin; Humans; Leukemia, Lymphoid; Leukemia, Myeloid; Leukemia, Myeloid, Acute; Lung Neoplasms; Male; Radioimmunoassay; Swine; Thrombosis

When Lewis lung carcinoma with low thromboplastic and low fibrinolytic activities was implanted subcutaneously to mice, administration of tranexamic acid inhibited metastasis formation in the lungs. This effect was considered to be mediated by prevention of cell release from the implanted sites. Fibrin formation around tumor cells in the vessels of primary foci was observed in the mice given tranexamic acid. On the other hand, urokinase significantly enhanced pulmonary metastases and many free tumor cells were observed intravascularly in primary foci of the mice given urokinase.

Topics: Animals; Cyclohexanecarboxylic Acids; Female; Fibrin; Lung Neoplasms; Male; Mice; Mice, Inbred C57BL; Mice, Inbred DBA; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms, Experimental; Rabbits; Tranexamic Acid; Urokinase-Type Plasminogen Activator

The antifibrinolytic action of tranexamic acid (AMCHA) on the growth and metastasis of rabbit V2 carcinoma having high fibrinolytic activity was studied. Upon oral administration of AMCHA, the growth of the tumor and metastasis to the lung tended to be inhibited, and the number of metastatic foci in the regional lymph nodes significantly decreased in the early period of tumor growth. Enhancement of fibrin deposition in the tumor and inhibition of fibrinolytic activity of the tumor were recognized in the AMCHA-treated group. The inhibitory effect of tranexamic acid on fibrin dissolution might interfere with local tumor growth and the release of tumor cells into the vessels.

Topics: Animals; Carcinoma; Cyclohexanecarboxylic Acids; Fibrin; Fibrinolysis; Lung Neoplasms; Lymphatic Metastasis; Male; Neoplasms, Experimental; Plasminogen Activators; Rabbits; Tranexamic Acid

Topics: Fibrin; Fibrinogen; Humans; Lung Neoplasms; Pleural Effusion; Tuberculosis, Pulmonary

Topics: Cells, Cultured; Endopeptidases; Fibrin; Lung Neoplasms; Neoplasms, Experimental; Proteoglycans

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

As shown in earlier studies the formation of metastases after i.v. tumor cell injection in rats is increased in the immediate post-traumatic period and treatment with heparin, thrombocytopenia, and defibrinogenation decreases the formation of metastases. Thrombocytopenia also inhibits the stimulating effect of trauma on metastasis formation. The results of the studies reported in this paper show that the changes of metastasis formation induced by the factors mentioned above with few exceptions are well correlated to the lodgement of the injected tumor cells. Thus, heparin treatment and thrombocytopenia decrease the pulmonary lodgement of i.v. injected tumor cells. Trauma increases the pulmonary lodgement of i.v. injected tumor cells but when trauma is combined with thrombocytopenia, the effect of thrombocytopenia dominates and the pulmonary lodgement of tumor cells decreases when compared to control conditions. Despite this, trauma still gives rise to increased tumor cell lodgement during thrombocytopenia.

Topics: Adenosine Diphosphate; Animals; Fibrin; Heparin; Lung Neoplasms; Neoplasm Metastasis; Neoplasms, Experimental; Neoplastic Cells, Circulating; Platelet Aggregation; Rats; Thrombocytopenia; Wounds and Injuries

Topics: Animals; Batroxobin; Fibrin; Humans; Lung Neoplasms; Mice; Neoplasm Metastasis; Neoplasm Recurrence, Local; Neoplasms, Experimental; Peptide Hydrolases; Time Factors

Effect of urokinase on the tumor growth and metastasis formation of rabbit V2 carcinoma, having low thromboplastic and high fibrinolytic activities, was examined. Weight of the tumor and lymphogenous metastases tended to increase, but the number of metastatic foci in the lungs was unchanged by the administration of urokinase. Diminution of fibrin deposits and connective tissue reaction in association with increase in the pattern of invasive growth was recognized at the advancing border of the tumors in the urokinase-treated group. In the intravenously induced pulmonary metastases, the number of metastatic foci decreased significantly in the urokinase-treated group. Fibrin was demonstrated at the site of tumor cell embolism by the conjugated anti-rabbit fibrinogen antibody. The growth of metastatic foci in the lungs was not affected by the treatment with urokinase. Enhancing effect of urokinase on fibrin resolution might promote the local tumor growth and release of tumor cells into the vessels, but interfere with the lodgement of tumor cells in remote organs.

Topics: Animals; Endopeptidases; Fibrin; Fibrinolysis; Lung Neoplasms; Male; Neoplasm Metastasis; Neoplasms, Experimental; Rabbits; Urokinase-Type Plasminogen Activator

Topics: Carcinoma, Bronchogenic; Diagnosis, Differential; Fibrin; Humans; Lung Neoplasms; Lymphoma; Mesothelioma; Neoplasm Metastasis; Pleural Diseases; Pleural Neoplasms; Radiography

Topics: Cell Transformation, Neoplastic; Cells, Cultured; Clot Retraction; Fibrin; Fibroblasts; Humans; Lung Neoplasms; Skin

Tumor emboli were produced in lungs of Sprague-Dawley rats by i.v. injection of Walker 256 tumor cells into the tail vein. Tissues were examined by electron microscopy at periods from 30 sec to 72 hr after tumor injection. Two methods of conventional staining were used, in addition to immunoperoxidase techniques, with antifibrin antibodies produced in rabbits. Tumor cells accompanied by a platelet mass were seen in pulmonary arterioles at the earliest time period (30 sec). By conventional staining, small amounts of fibrin were detected within the platelet clumps by 5 min after inoculation. Periodicity indicating stable fibrin was not seen by this technique until 15 to 45 min. When peroxidase-labeled antibody was applied to tissue, sections showed fibrin-positive material at 30 sec, and periodicity of fibrin was detected by 5 min. Fibrin reached a maximum by both techniques at about 1 hr and disappeared, along with the platelets, at about 9 hr. When fibrinolysin was injected prior to the tumor cell inoculation, platelets and fibrin were either absent or present only in traces, and no stable fibrin was detected. These observations show that fibrin occurs very early in small amounts in association with tumor cell emboli, and is removed while the cells are still intravascular.

Topics: Animals; Blood Platelets; Carcinoma 256, Walker; Fibrin; Lung Neoplasms; Neoplasm Metastasis; Platelet Aggregation; Rats; Time Factors

Topics: Adenocarcinoma; Adult; Aged; Alpha-Globulins; Blood Coagulation Disorders; Blood Protein Electrophoresis; Bronchial Neoplasms; Carcinoma; Carcinoma, Bronchogenic; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Fibrin; Fibrinogen; Fibrinolysin; Fibrinolysis; Heparin; Humans; Infusions, Parenteral; Lung Neoplasms; Macroglobulins; Male; Middle Aged; Time Factors

Topics: Aminocaproates; Blood Coagulation; Breast Neoplasms; Colonic Neoplasms; Female; Fibrin; Fibrinogen; Fibrinolysis; Gastrointestinal Neoplasms; Humans; Immunoelectrophoresis; Intestinal Neoplasms; Kidney Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Male; Neoplasm Metastasis; Neoplasm Recurrence, Local; Pancreatic Neoplasms; Prostatic Neoplasms; Rectal Neoplasms; Thrombin; Urinary Bladder Neoplasms; Urogenital Neoplasms

Topics: Animals; Blood Platelets; Capillaries; Carcinoma 256, Walker; Cell Aggregation; Cell Count; Fibrin; Fluorescent Antibody Technique; Goats; Histocytochemistry; Immune Sera; Immunodiffusion; Lung Neoplasms; Male; Methods; Microscopy, Electron; Neoplasm Metastasis; Neoplasms, Experimental; Pulmonary Artery; Rabbits; Rats; Staining and Labeling; Time Factors

Topics: Aminocaproates; Aminocaproic Acid; Animals; Aprotinin; Blood Coagulation; Carcinoma 256, Walker; Carcinoma, Brown-Pearce; Fibrin; Fibrinolysin; Heparin; Hyperlipidemias; Lung Neoplasms; Neoplasm Metastasis; Neoplasm Transplantation; Neoplasms; Neoplasms, Experimental; Nitrogen Mustard Compounds; Pharmacology; Rabbits; Radiation Effects; Rats; Research

Topics: Animals; Blood Coagulation; Brain Neoplasms; Carbon Tetrachloride Poisoning; Carcinoma 256, Walker; Female; Fibrin; Heart Neoplasms; In Vitro Techniques; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Male; Mesentery; Neoplasm Metastasis; Neoplastic Cells, Circulating; Rats; Testicular Neoplasms; Wounds and Injuries

Topics: Carcinoma; Carcinoma, Bronchogenic; Diagnosis, Differential; Fibrin; Humans; Lung Neoplasms

Topics: Fibrin; Fibrinolysis; Humans; Lung Neoplasms

Topics: Abscess; Fibrin; Humans; Lung; Lung Abscess; Lung Neoplasms; Neoplasms

Topics: Carcinoma, Bronchogenic; Diagnosis, Differential; Fibrin; Humans; Lung Neoplasms