fibrin and Breast-Neoplasms

Fibrin sheath formation around venous access devices (VADs) frequently leads to persistent withdrawal occlusion (PWO). PWO is a common problem encountered with VADs. Although PWO is often easily managed with small doses of thrombolytic therapy (e.g., urokinase), it could result in a more serious complication, such as chemotherapy extravasation. Careful assessment of all VADs is important to identify complications such as fibrin sheath formation, which can potentially lead to extravasation. To rule out fibrin sheath formation, catheter dye studies need to be obtained when fibrinolytic therapy has failed to restore catheter function. The purpose of this paper is to illustrate a retrospective case report demonstrating drug extravasation caused by the development of fibrin sheath formation.

Topics: Antineoplastic Combined Chemotherapy Protocols; Breast Neoplasms; Catheterization, Central Venous; Catheters, Indwelling; Constriction, Pathologic; Cyclophosphamide; Equipment Failure; Extravasation of Diagnostic and Therapeutic Materials; Female; Fibrin; Humans; Infusions, Intravenous; Middle Aged; Paclitaxel; Plasminogen Activators; Urokinase-Type Plasminogen Activator

Topics: Absorption; Amylases; Antineoplastic Agents; Breast Neoplasms; Bronchial Neoplasms; Capillary Permeability; Carcinoembryonic Antigen; Catheterization; Drainage; Dyspnea; Fibrin; Filtration; Humans; Hydrogen-Ion Concentration; Lymphoma; Pain; Pancreatic Neoplasms; Pleural Effusion; Pleural Neoplasms; Pregnancy Proteins; Quinacrine; Talc; Tetracyclines; Tissue Adhesives; Ultrasonography

Topics: Adult; Antineoplastic Agents; Blood Coagulation; Blood Coagulation Factors; Breast Neoplasms; Dextrans; Female; Fibrin; Fibrinogen; Fibrinolysis; Humans; Male; Middle Aged; Neoplasm Metastasis; Neoplasms; Ovarian Neoplasms; Rectal Neoplasms; Sulfates

The progression of breast cancer involves cancer-cell invasions of extracellular matrices. To investigate the progression, 3D cell cultures are widely used along with different types of matrices. Currently, the matrices are often characterized using parallel-plate rheometry for matrix viscoelasticity, or liquid-like viscous and stiffness-related elastic characteristics. The characterization reveals averaged information and sample-to-sample variation, yet, it neglects internal heterogeneity within matrices, experienced by cancer cells in 3D culture. Techniques using optical tweezers and magnetic microrheometry have measured heterogeneity in viscoelasticity in 3D culture. However, there is a lack of probabilistic heterogeneity quantification and cell-size-relevant, microscale-viscoelasticity measurements at breast-tumor tissue stiffness up to ≃10 kPa in Young's modulus. Here, we have advanced methods, for the purpose, which use a magnetic microrheometer that applies forces on magnetic spheres within matrices, and detects the spheres displacements. We present probabilistic heterogeneity quantification using microscale-viscoelasticity measurements in 3D culture matrices at breast-tumor-relevant stiffness levels. Bayesian multilevel modeling was employed to distinguish heterogeneity in viscoelasticity from the effects of experimental design and measurement errors. We report about the heterogeneity of breast-tumor-relevant agarose, GrowDex, GrowDex-collagen and fibrin matrices. The degree of heterogeneity differs for stiffness, and phase angle (i.e. ratio between viscous and elastic characteristics). Concerning stiffness, agarose and GrowDex show the lowest and highest heterogeneity, respectively. Concerning phase angle, fibrin and GrowDex-collagen present the lowest and the highest heterogeneity, respectively. While this heterogeneity information involves softer matrices, probed by ≃30 μm magnetic spheres, we employ larger ≃100 μm spheres to increase magnetic forces and acquire a sufficient displacement signal-to-noise ratio in stiffer matrices. Thus, we show pointwise microscale viscoelasticity measurements within agarose matrices up to Young's moduli of 10 kPa. These results establish methods that combine magnetic microrheometry and Bayesian multilevel modeling for enhanced heterogeneity analysis within 3D culture matrices.

Topics: Bayes Theorem; Breast Neoplasms; Cell Culture Techniques, Three Dimensional; Collagen; Elastic Modulus; Extracellular Matrix; Female; Fibrin; Humans; Magnetic Phenomena; Sepharose

Topics: Breast Neoplasms; Cytokines; Female; Fibrin; Granzymes; Humans; Interferon-gamma; Interleukin-13; Ligands; Minor Histocompatibility Antigens; Mucosal-Associated Invariant T Cells; Receptors, Antigen, T-Cell; T-Lymphocytes; Tumor Microenvironment

Topics: Breast Implants; Breast Neoplasms; Diagnosis, Differential; Female; Fibrin; Humans; Lymphoma, Large B-Cell, Diffuse; Lymphoma, Large-Cell, Anaplastic; Middle Aged

Metastasis remains the major cause of death in cancer patients. Thus, there is a need to sensitively detect tumor metastasis, especially ultrasmall metastasis, for early diagnosis and precise treatment of cancer. Herein, an ultrasensitive T

Topics: Animals; Breast Neoplasms; Cell Line, Tumor; Contrast Media; Female; Ferric Compounds; Fibrin; Fibronectins; Humans; Hydrogen Peroxide; Magnetic Resonance Imaging; Manganese Compounds; Mice; Mice, Inbred BALB C; Nanoparticles; Neoplasm Metastasis; Oligopeptides; Signal-To-Noise Ratio; Transplantation, Heterologous

Secretion of PDI from platelets and endothelial cells is an important step of all thrombotic events. In the absence of extracellular PDI thrombus formation and fibrin generation may be impaired. Thrombin-mediated PDI secretion is regulated by the stimulation of P2Y

Topics: Androgen Receptor Antagonists; Blood Platelets; Blood Specimen Collection; Bodily Secretions; Breast Neoplasms; Cell Line, Tumor; Cell Movement; Endothelial Cells; Endothelium; Female; Fibrin; Humans; Neoplasm Metastasis; Platelet Adhesiveness; Protein Disulfide-Isomerases; Purinergic P2Y Receptor Antagonists; Receptors, Androgen; Receptors, Purinergic P2Y12; Signal Transduction; Sulfhydryl Compounds; Thrombin; Thrombosis

Molecular crosstalk between intra-tumor blood vessels and tumor cells plays many critical roles in tumorigenesis and cancer metastasis. However, it has been very difficult to investigate the biochemical mechanisms underlying the overlapping, multifactorial processes that occur at the tumor-vascular interface using conventional murine models alone. Moreover, traditional two-dimensional (2D) culture models used in cancer research do not recapitulate aspects of the 3D tumor microenvironment. In the present study, we introduce a microfluidic model of the solid tumor-vascular interface composed of a human umbilical vein endothelial cell (HUVEC)-lined, perfusable, bioengineered blood vessel and tumor spheroids embedded in an extracellular matrix (ECM). We sought to optimize our model by varying the composition of the tumor spheroids (MDA-MB-231 breast tumor cells + mesenchymal stem cells (MSCs)/human lung fibroblasts (HLFs)/HUVECs) and the extracellular matrix (ECM: collagen, Matrigel, and fibrin gels with or without free HLFs) that we used. Our results indicate that culturing tumor spheroids containing MDA-MB-231 cells + HUVECs in an HLF-laden, fibrin-based ECM within our microfluidic device optimally (1) enhances the sprouting and migration of tumor spheroids, (2) promotes angiogenesis, (3) facilitates vascular invasion, and (4) preserves the structural integrity and functionality of HUVEC-lined microfluidic channels. This model may provide a platform for drug screening and mechanism studies on solid tumor interactions with functional blood vessels.

Topics: Blood Vessels; Breast Neoplasms; Cell Line, Tumor; Collagen; Drug Combinations; Extracellular Matrix; Fibrin; Human Umbilical Vein Endothelial Cells; Humans; Lab-On-A-Chip Devices; Laminin; Mesenchymal Stem Cells; Neovascularization, Pathologic; Perfusion; Proteoglycans; Spheroids, Cellular; Tissue Culture Techniques; Tumor Microenvironment

We examined the link between gestational biomarkers and breast cancer in 9169 daughters born into the Child Health and Development Studies from 1959 to 1967. We identified 137 breast cancer cases diagnosed by age 52 as of 2012. Markers of increased risk included higher placental volume and rapid 2nd trimester gestational weight gain. Protective markers were placental hemorrhage and fibrin deposition, indicators of resistance to placental trophoblast invasion. Paradoxically, higher ponderal index at birth was protective suggesting that fetal and placental pathways to breast cancer are multiple and distinct. Results link placental and fetal phenotypes to breast cancer, characterizing some as restrictive and others as permissive markers of tumor development. We found new biomarkers of breast cancer risk that can be mined to discover 'omic correlates in the pregnancy exposome using archived and contemporary pregnancy samples. This line of investigation may discover new pathways to risk and new opportunities for prevention.

Topics: Biomarkers; Breast Neoplasms; California; Child; Child Development; Child Health; Female; Fetal Development; Fibrin; Hemorrhage; Humans; Nuclear Family; Phenotype; Placenta; Pregnancy; Pregnancy Trimester, Second; Risk Factors; Weight Gain

Tumor cell-induced platelet aggregation represents a critical process both for successful metastatic spread of the tumor and for the development of thrombotic complications in cancer patients. To get further insights into this process, we investigated and compared the molecular mechanisms of platelet aggregation induced by two different breast cancer cell lines (MDA-MB-231 and MCF7) and a colorectal cancer cell line (Caco-2). All the three types of cancer cells were able to induce comparable platelet aggregation, which, however, was observed exclusively in the presence of CaCl

Topics: Amino Acid Chloromethyl Ketones; Aspirin; Blood Platelets; Breast Neoplasms; Caco-2 Cells; Calcium Chloride; Colorectal Neoplasms; Female; Fibrin; Fibrinogen; Humans; Integrin alpha2; MCF-7 Cells; Platelet Activation; Platelet Aggregation; Thromboxane A2; Type C Phospholipases

Cancer progression is associated with an evolving tissue interface of direct epithelial-tumor microenvironment interactions. In biopsies of human breast tumors, extensive alterations in molecular pathways are correlated with cancer staging on both sides of the tumor-stroma interface. These interactions provide a pivotal paracrine signaling to induce malignant phenotype transition, the epithelial-mesenchymal transition (EMT). We explored how the direct contact between platelets-fibrin bundles primes metastasis using platelet-rich plasma (PRP) as a source of growth factors and mimics the provisional fibrin matrix between actively growing breast cancer cells and the tumor stroma. We have demonstrated PRP functions, modulating cell proliferation that is tumor-subtype and cancer cell-type-specific. Epithelial and stromal primary cells were prepared from breast cancer biopsies from 21 women with different cancer subtypes. Cells supplemented with PRP were immunoblotted with anti-phospho and total Src-Tyr-416, FAK-Try-925, E-cadherin, N-cadherin, TGF-β, Smad2, and Snail monoclonal antibodies. Breast tumor cells from luminal B and HER2 subtypes showed the most malignant profiles and the expression of thrombin and other classes of proteases at levels that were detectable through FRET peptide libraries. The angiogenesis process was investigated in the interface obtained between platelet-fibrin-breast tumor cells co-cultured with HUVEC cells. Luminal B and HER2 cells showed robust endothelial cell capillary-like tubes ex vivo. The studied interface contributes to the attachment of endothelial cells, provides a source of growth factors, and is a solid substrate. Thus, replacement of FBS supplementation with PRP supplementation represents an efficient and simple approach for mimicking the real multifactorial tumor microenvironment.

Topics: Blood Platelets; Breast Neoplasms; Disease Progression; Epithelial Cells; Female; Fibrin; Human Umbilical Vein Endothelial Cells; Humans; Neovascularization, Pathologic; Platelet-Rich Plasma; Stromal Cells; Tumor Microenvironment

Polymyalgia rheumatica (PMR), a chronic inflammatory rheumatism, can be the expression of a paraneoplastic syndrome. The same clinical symptoms are frequently observed at the early stage of the benign and malignant forms. Here, our aim was to develop diagnostic tools to differentiate paraneoplastic PMR from essential PMR. We combined an 18FDG-PET and detection of circulating procoagulant microparticles (MPs), such as fibrin positive (FibMPs), by flow cytometry. Two patients with PMR and a similar profile were selected. In the two patients, the 18FDG-PET revealed a hypermetabolic focus. However, the concentrations of fibrin+/annexin+ microparticles detected were (10 times higher in one of the two patients, who was later found to have breast cancer. The association of 18FDG-PET and the detection of microparticle fibrin positives by flow cytometry allows separating essential PMR (hypermetabolism by 18FDG-PET, low FibMPs) from paraneoplastic PMR.

Topics: Aged; Breast Neoplasms; Cell-Derived Microparticles; Colonic Neoplasms; Female; Fibrin; Fluorodeoxyglucose F18; Humans; Middle Aged; Paraneoplastic Syndromes; Polymyalgia Rheumatica; Positron Emission Tomography Computed Tomography; Whole Body Imaging

Thromboembolic complications are a common cause of death in breast cancer patients. The in vivo relationship between coagulation factors and circulating tumours is a multifaceted one, with tumour cells implicated in thrombocytosis and platelets associated with coagulation-mediated metastasis. Platelets and fibrin networks are known to be morphologically altered in patients with cancer, and their relationship with breast cancer cells themselves may increase thrombosis susceptibility. This was investigated in vitro, assessing such morphological alterations through the establishment of a MCF-7 breast cancer cell co-culture system with blood plasma. Co-culture duration ranged from zero to thirty minutes, with five-minute intervals, in order to assess the time-dependent ultrastructural conformations of platelet and fibrin networks, using scanning electron microscopy. It was found that enhanced coagulability was related to co-culture duration. Changes in platelet and fibrin network morphology from normal were visible as early as five minutes in co-culture with MCF-7 cells. With longer co-culture duration thrombosis-linked variation in structural configuration was intensified, including advanced platelet aggregation and adherence characteristics, compression of fibrin networks into plaques of increased density, and upsurge of microparticulate extrusion implicated in amplifying procoagulant events during the metastatic process. These results confirm that cancer cells are stimulators of coagulation even in an in vitro system and breast cancer patients may become increasingly susceptible to thrombotic-related consequences with increased exposure to tumour cells, especially during metastasis.

Topics: Blood Coagulation; Blood Platelets; Breast Neoplasms; Coculture Techniques; Female; Fibrin; Humans; MCF-7 Cells; Microscopy, Electron, Scanning; Thromboembolism; Thrombosis; Time Factors

Migration stimulating factor (MSF) is an oncofetal motogenic/angiogenic cytokine constitutively expressed by epithelial and stromal cells in fetal and neoplastic tissues. Fibroblasts derived from healthy adult skin do not express MSF but can be induced to do so by treatment with transforming growth factor-β1 (TGF-β1). As the bioactivities of both MSF and TGF-β1 are modulated by the extracellular matrix, we investigated whether the induction of MSF expression by TGF-β1 is also matrix dependent. We now report that adult fibroblasts are induced to express MSF by a transient treatment with TGF-β1 (as short as 2 hr) but only when the cells are adherent to a "wound" matrix, such as denatured type I collagen, fibrin or plastic tissue culture dishes. Unexpectedly, this induction of MSF expression persists unabated for the entire subsequent lifespan of the treated cells in the absence of further TGF-β1 and irrespective of the substratum. Such "activated" MSF expression may, however, be persistently switched off again by a second transient exposure to TGF-β1 but this time only when the cells are adherent to a "healthy" matrix of native type I collagen. Significantly, the constitutive expression of MSF by fetal and cancer patient fibroblasts could also be persistently switched off by this means. We conclude that TGF-β1 may both switch on and switch off MSF expression in a manner critically determined by the nature of the matrix substratum and suggest that this may be a possible mechanism underlying the observed dual functionality of TGF-β1 as both a tumour suppressor and tumour promoter.

Topics: Animals; Breast Neoplasms; Carcinoma, Ductal; Cell Migration Assays; Cells, Cultured; Collagen Type I; Cytokines; Extracellular Matrix; Female; Fetal Development; Fibrin; Fibroblasts; Fibronectins; Gene Expression Regulation; Humans; Middle Aged; Protein Isoforms; Rats; Signal Transduction; Transforming Growth Factor beta1

Fibrin deposition has been indicated within the stroma of a majority of solid tumors. Here we assess the feasibility of using the established fibrin-specific probe EP-2104R for noninvasive imaging of fibrin in the context of breast cancer.. EP-2104R, untargeted gadopentetate dimeglumine (Gd-DTPA), and a newly synthesized nonfibrin binding control linear peptide (CLP) were compared using steady-state and dynamic contrast-enhanced magnetic resonance imaging in a breast cancer xenograft mouse model at 9.4 T.. EP-2104R transiently enhanced both tumor core and tumor periphery, but only the enhancement in the tumor periphery persisted even 90 minutes after EP-2104R administration. However, untargeted Gd-DTPA and CLP are not retained in the tumor periphery. The half-life of EP-2104R in the tumor periphery (103 ± 18 minutes) is significantly longer (P < 0.05) than that of either Gd-DTPA (29.6 ± 2.4 minutes) or CLP (42.4 ± 1.5 minutes), but the rate of clearance is similar for all the 3 probes from the tumor core. The presence of high concentrations of fibrin in the tumor periphery was corroborated using immunohistochemistry with a fibrin-specific antibody.. The persistent enhancement observed in the tumor periphery with EP-2104R is likely a result of its fibrin-specific binding rather than its size and demonstrates the feasibility of EP-2104R for molecular imaging of fibrin in tumor stroma.

Topics: Adenocarcinoma; Animals; Breast Neoplasms; Disease Models, Animal; Feasibility Studies; Female; Fibrin; Gadolinium; Gadolinium DTPA; Mice; Molecular Imaging; Peptides; Transplantation, Heterologous

The aim of our study was to explain the effect of elevated homocysteine (measured by HPLC) on haemostatic activity of plasma from breast cancer patients (fibrin polymerization and lysis; the thrombin and prothrombin time), because homocysteine (Hcys) induces changes in haemostasis, as well blood clotting as fibrinolysis. Patients were hospitalized in Department of Oncological Surgery, Medical University of Lodz, Poland. All patients have not had preadjuvant therapy, and samples from patients were taken before surgery. We observed that changes of selected parameters of haemostatic properties of plasma, e.g., the prothrombin time and thrombin time were prolonged in plasma from invasive breast cancer when compared with the control group (healthy subjects) and patients with benign breast diseases. Our results showed also that the correlation between the increased amount of Hcys and changes of selected parameters of haemostasis in invasive breast cancer patients exists. Considering the data presented in this study, we suggest that the elevated Hcys in invasive breast cancer patients may induce the changes of haemostatic properties of plasma isolated from these patients.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Carcinoma, Ductal, Breast; Case-Control Studies; Female; Fibrin; Fibrinolysis; Hemostasis; Homocysteine; Humans; Middle Aged; Risk Factors; Statistics, Nonparametric; Thrombin Time

Many cytotoxic drugs maintain antiangiogenic properties, but there are no human, tumor-based assays to evaluate their antiangiogenic potential. We used a fibrin-thrombin clot-based angiogenesis model to evaluate the angiogenic response of human breast cancer to various cytotoxic agents commonly used in its treatment.. Fragments of freshly harvested human breast tumors were embedded in fibrin-thrombin clots and treated with five drugs: adriamycin, taxol, 5-fluorouracil (5-FU), methotrexate, and vincristine. Each treatment group included a mean of 28 fragments (range 16-60). A total of four tumors were tested. Tumor fragments were tested with a single dose of each reagent. Angiogenic initiation, angiogenic growth, and overall angiogenic effect were determined for each treatment group using a previously validated scale.. All four breast cancer specimens tested developed an angiogenic response, sprouting neovessels in vitro in a time-dependent fashion (r = 0.84, P = 0.0007). Taxol statistically inhibited angiogenesis in all four specimens with decreases in the mean angiogenic initiation, angiogenic growth, and overall effect that were 69%, 81%, and 94% of control values, respectively. Vincristine and 5-FU inhibited the mean overall angiogenic effect by 89% and 82% compared with control, respectively. Adriamycin inhibited overall effect 49%. Methotrexate was less effective.. Freshly harvested breast cancer specimens develop an angiogenic response in a fibrin-thrombin clot-based angiogenesis model and respond to treatment with antineoplastic/antiangiogenic drugs. The antiangiogenic potential of commonly used breast cancer drugs varied among individual tumors. Data obtained from this model is unique and might potentially be used to further enhance the efficacy of cytotoxic regimens and individualize patient therapy.

Topics: Angiogenesis Inhibitors; Antibiotics, Antineoplastic; Antimetabolites, Antineoplastic; Antineoplastic Agents, Phytogenic; Biological Assay; Blood Coagulation; Breast Neoplasms; Doxorubicin; Drug Screening Assays, Antitumor; Female; Fibrin; Fluorouracil; Humans; Methotrexate; Neoplasm Invasiveness; Neovascularization, Pathologic; Paclitaxel; Thrombin; Vincristine

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

Recent evidence assigns integrins and metalloproteinases (MMPs) an important role in regulating tumor cell progression. Here, we demonstrate that 3-O-methylfunicone (OMF), a secondary metabolite produced by Penicillium pinophilum, affects cell proliferation and motility of breast cancer MCF-7 cells, downregulating alphavbeta5 integrin, and inhibiting MMP-9 secretion. This effect was absent when the non-tumoral MCF-10 cell line was used. Inhibition of cell motility was also associated to modifications in cell shape and in the distribution of tubulin fibers of OMF-treated MCF-7 cells. In addition, a possible effect on survivin and hTERT was also investigated. We found that OMF strongly inhibits survivin and hTERT gene expression. The results of this study indicate that OMF-induced inhibition of cell motility may be mediated through the modulation of alphavbeta5 integrin and MMP-9 secretion. In addition, the inhibition of typical markers of tumor progression such as hTERT and survivin in MCF-7 and their inactivity towards MCF10 provide strong evidence for a potential use of OMF in anticancer therapy.

Topics: Breast Neoplasms; Cell Line, Tumor; Cell Movement; Cell Proliferation; Cell Shape; Down-Regulation; Drug Evaluation; Female; Fibrin; Humans; Integrins; Matrix Metalloproteinase 9; Penicillium; Pyrones; Receptors, Vitronectin; Tubulin

Although angiogenesis is a prerequisite for the growth of most human solid tumours, alternative mechanisms of vascularisation can be adopted. We have previously described a non-angiogenic growth pattern in liver metastases of colorectal adenocarcinomas (CRC) in which tumour cells replace hepatocytes at the tumour-liver interface, preserving the liver architecture and co-opting the sinusoidal blood vessels. The aim of this study was to determine whether this replacement pattern occurs during liver metastasis of breast adenocarcinomas (BC) and whether the lack of an angiogenic switch in such metastases is due to the absence of hypoxia and subsequent vascular fibrinogen leakage. The growth pattern of 45 BC liver metastases and 28 CRC liver metastases (73 consecutive patients) was assessed on haematoxylin- and eosin-stained tissue sections. The majority of the BC liver metastases had a replacement growth pattern (96%), in contrast to only 32% of the CRC metastases (P<0.0001). The median carbonic anhydrase 9 (CA9) expression (M75 antibody), as a marker of hypoxia, (intensity x % of stained tumour cells) was 0 in the BC metastases and 53 in the CRC metastases (P<0.0001). There was CA9 expression at the tumour-liver interface in only 16% of the BC liver metastases vs 54% of the CRC metastases (P=0.002). There was fibrin (T2G1 antibody) at the tumour-liver interface in only 21% of the BC metastases vs 56% of the CRC metastases (P=0.04). The median macrophage count (Chalkley morphometry; KP-1 anti-CD68 antibody) at the interface was 4.3 and 7.5, respectively (P<0.0001). Carbonic anhydrase 9 score and macrophage count were positively correlated (r=0.42; P=0.002) in all metastases. Glandular differentiation was less in the BC liver metastases: 80% had less than 10% gland formation vs only 7% of the CRC metastases (P<0.0001). The liver is a densely vascularised organ and can host metastases that exploit this environment by replacing the hepatocytes and co-opting the vasculature. Our findings confirm that a non-angiogenic pattern of liver metastasis indeed occurs in BC, that this pattern of replacement growth is even more prevalent than in CRC, and that the process induces neither hypoxia nor vascular leakage.

Topics: Adenocarcinoma; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Breast Neoplasms; Carbonic Anhydrases; Cell Hypoxia; Colorectal Neoplasms; Fibrin; Glycoproteins; Humans; Immunohistochemistry; Liver Neoplasms; Macrophages; Neovascularization, Pathologic; Vesicular Transport Proteins

This study investigated the connection among HER-2 gene amplification, HER-2 protein expression, and markers of tumor angiogenesis and oxygenation in patients with operable, invasive breast tumors.. From 1988 to 1995, 425 patients with metastatic breast cancer were enrolled in a study of high-dose chemotherapy with autologous transplant. Primary tumor blocks were obtained and evaluated using immunohistochemistry (IHC) staining of vessels with von Willebrand factor antibody. Mean microvessel densities (MVD) were determined by counting von Willebrand factor stained cells in three separate "vascular hot spots" using image analysis. Tumor samples were also stained for HER-2 by IHC, HER-2 gene amplification by fluorescence in situ hybridization, carbonic anhydrase 9 by IHC, and vascular endothelial growth factor (VEGF) by IHC. Plasma from 36 patients with primary tumor samples had VEGF (R&D Systems, MN) and d-dimer (American Diagnostica, Greenwich, CT) levels determined.. There was a significant positive correlation between HER-2 gene amplification and both maximum and average MVD (Spearman coefficient = 0.51 and 0.50; P = 0.03 and 0.05, respectively). There was an inverse correlation with HER-2 gene amplification and expression of the tumor hypoxia marker CA-9 (chi(2) P = 0.02). The level of HER-2 gene amplification correlated with plasma d-dimer levels (Spearman coefficient = 0.43; P = 0.021). Interestingly, tumors with HER-2 by IHC had decreased amounts of VEGF staining (chi(2) = 5.81; P = 0.01). There was no correlation between HER-2 by IHC and MVD or d-dimer. Of all of the variables examined, only average (P = 0.0016) and maximum MVD (P = 0.0128) predicted disease-free survival (Cox univariate model).. HER-2-amplified breast cancers have increased amounts of angiogenesis, decreased amounts of hypoxia, and increased markers of fibrin degradation. These findings have prognostic, predictive, and therapeutic implications in breast cancer treatment.

Topics: Breast Neoplasms; Carbonic Anhydrases; Disease-Free Survival; Enzyme-Linked Immunosorbent Assay; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Humans; Hypoxia; Immunohistochemistry; In Situ Hybridization, Fluorescence; Microcirculation; Neoplasms; Neovascularization, Pathologic; Oxygen; Prognosis; Receptor, ErbB-2; Time Factors; Vascular Endothelial Growth Factor A; von Willebrand Factor

Inflammatory breast cancer (IBC) is the most aggressive form of breast cancer. Improved understanding of the mechanisms responsible for the differences between IBC and non-IBC might provide novel therapeutic targets. We studied 35 consecutive patients with IBC, biopsied prior to the initiation of chemotherapy. Angiogenesis was evaluated by Chalkley counting and by assessment of endothelial cell proliferation (ECP) and vessel maturity. The presence of fibrin, expression of the hypoxia marker carbonic anhydrase IX (CA IX) and epithelialcadherin (E-cadherin) expression were immunohistochemically detected. The same parameters were obtained in a group of 104 non-IBC patients. Vascular density, assessed by Chalkley counting (P<0.0001), and ECP (P=0.01) were significantly higher in IBC than in non-IBC. Abundant stromal fibrin deposition was observed in 26% of IBC and in only 8% of non-IBC (P=0.02). Expression of CA IX was significantly less frequent in IBC than in non-IBC with early metastasis (P=0.047). There was a significant positive correlation between the expression of CA IX and ECP in IBC (r=0.4, P=0.03), implying that the angiogenesis is partly hypoxia driven. However, the higher ECP in IBC and the less frequent expression of CA IX in IBC vs non-IBC points at a role for other factors than hypoxia in stimulating angiogenesis. Strong, homogeneous E-cadherin expression was found at cell-cell contacts in all but two IBC cases, both in lymphovascular tumour emboli and in infiltrating tumour cells, challenging our current understanding of the metastatic process. Both the intense angiogenesis and the strong E-cadherin expression may contribute to the highly metastatic phenotype of IBC.

Topics: Adult; Aged; Aged, 80 and over; Breast Neoplasms; Cadherins; Carbonic Anhydrases; Cell Division; Endothelium, Vascular; Fibrin; Humans; Middle Aged; Neovascularization, Pathologic

Localization of plasmin activity on leukocyte surfaces plays a critical role in fibrinolysis as well as in pathological and physiological processes in which cells must degrade the extracellular matrix in order to migrate. The binding of plasminogen to leukocytic cell lines induces a 30- to 80-fold increase in the rate of plasminogen activation by tissue-type (tPA) and urokinase-type (uPA) plasminogen activators. In the present study we have examined the role of alpha-enolase in plasminogen activation on the cell surface. We produced and characterized a monoclonal antibody (MAb) 11G1 against purified alpha-enolase, which abrogated about 90% of cell-dependent plasminogen activation by either uPA or tPA on leukocytoid cell lines of different lineages: B-lymphocytic, T-lymphocytic, granulocytic, and monocytic cells. In addition, MAb 11G1 also blocked enhancement of plasmin formation by peripheral blood neutrophils and monocytes. In contrast, MAb 11G1 did not affect plasmin generation in the presence of fibrin, indicating that this antibody did not interact with fibrinolytic components in the absence of cells. These data suggest that, although leukocytic cells display several molecules that bind plasminogen, alpha-enolase is responsible for the majority of the promotion of plasminogen activation on the surfaces of leukocytic cells.

Topics: Adenocarcinoma; Antibodies, Monoclonal; B-Lymphocytes; Blood Cells; Breast Neoplasms; Carboxypeptidase B; Carboxypeptidases; Depression, Chemical; Female; Fibrin; Fibrinogen; Fibrinolysin; Fibrinolysis; Humans; Leukocytes; Neoplasm Invasiveness; Neoplasm Proteins; Peptide Fragments; Phosphopyruvate Hydratase; Plasminogen; Protein Binding; Subcellular Fractions; Thrombin; Tissue Plasminogen Activator; Tumor Cells, Cultured; Urokinase-Type Plasminogen Activator

We postulated that skin metastases and cutaneous local recurrences from breast adenocarcinoma show different growth patterns with distinct angiogenic profiles.. Fifty-one surgically resected dermal breast cancer deposits were evaluated for growth pattern, E-cadherin expression, presence of necrosis and a fibrotic focus, fibrin deposition, carbonic anhydrase IX expression (CA IX), microvessel density, endothelial cell proliferation and blood vessel immaturity. Growth patterns were infiltrative, with carcinoma cells infiltrating the dermis without significant disturbance of the pre-existing architecture, expansive, meaning that a nodule of carcinoma cells and desmoplastic tissue pushed aside the pre-existing dermal structures, or mixed. All lobular carcinomas showed an infiltrative growth and lacked membranous E-cadherin expression. Different growth patterns in the ductal carcinomas were not correlated with differences in E-cadherin expression. The presence of necrosis and/or a fibrotic focus and the expression of the hypoxia marker CA IX were significantly associated with an expansive growth. Fibrin was present in all expansive deposits and less frequently in the other growth patterns. There was a positive association between fibrin deposition, CA IX expression and microvessel density. The latter was significantly higher in the expansive and mixed growth patterns than in the infiltrative pattern. Endothelial cell proliferation was highest in the expansive growth pattern and was positively correlated with the presence of a fibrotic focus and with fibrin deposition. The maximum percentage of immature blood vessels was higher in the expansive and mixed growth patterns than in the infiltrative one.. The recognition of different subgroups of cutaneous breast cancer deposits with different degrees of hypoxia-driven angiogenesis may have important implications for the usefulness of anti-angiogenic therapy.

Topics: Adenocarcinoma; Antigens, Neoplasm; Biomarkers, Tumor; Breast Neoplasms; Cadherins; Carbonic Anhydrase IX; Carbonic Anhydrases; Cell Hypoxia; Female; Fibrin; Fluorescent Antibody Technique, Indirect; Humans; Immunoenzyme Techniques; Microcirculation; Necrosis; Neoplasm Proteins; Neovascularization, Pathologic; Skin; Skin Neoplasms

Topics: Blood Coagulation Disorders; Breast Neoplasms; Cell Hypoxia; Female; Fibrin; Fibrinogen; Humans; Middle Aged; Neoplasm Metastasis

Seven patients with mammographic lesions entirely removed at percutaneous core needle biopsy that required wider excision underwent freehand localization of the site of the prior lesion with orthogonal and reproducible mammographic landmarks to guide needle placement. Successful excision was accomplished in all cases, as evidenced by similar histopathologic findings, fibrin bands or collagen, and core needle biopsy tract at microscopy.

Topics: Aged; Biopsy, Needle; Breast; Breast Neoplasms; Carcinoma in Situ; Carcinoma, Ductal, Breast; Collagen; Coloring Agents; Feasibility Studies; Female; Fibrin; Follow-Up Studies; Humans; Hyperplasia; Mammography; Methylene Blue; Middle Aged; Needles; Neoplasm, Residual; Preoperative Care; Radiography, Interventional; Stereotaxic Techniques

Topics: Antigens, CD; Antigens, CD34; Blood Cells; Bone Marrow Transplantation; Breast Neoplasms; Cells, Cultured; Colony-Forming Units Assay; Female; Fibrin; Hematopoietic Stem Cell Transplantation; Hematopoietic Stem Cells; Humans; In Vitro Techniques; Leukapheresis; Megakaryocytes; Methylcellulose; Thrombocytopenia; Transplantation, Autologous

This study was undertaken to determine if primary breast tumor plasminogen activator expression correlates with skeletal metastasis in breast cancer. Total plasminogen activator activity was significantly lower in tumors of patients with recurrence than in recurrence-free patients. Similarly, the primary tumors of patients with skeletal metastasis contained considerably less enzyme activity compared with those of patients surviving without skeletal metastasis. When patients with skeletal metastasis were categorized in terms of their recurrence pattern, those who had skeletal metastasis without other organ metastasis had significantly less tissue-type plasminogen activator antigen in their primary breast tumors than did those who had metastasis to other organs. Furthermore, a significantly lower level of tissue-type plasminogen activator antigen was found in primary tumors associated with axial bone metastasis than in those associated with appendicular bone metastasis. These results suggest that tissue-type plasminogen activator is involved in skeletal metastasis formation by its effects through the vertebral venous plexus.

Topics: Bone Neoplasms; Breast Neoplasms; Fibrin; Fibrinolysis; Humans; Neoplasm Metastasis; Neoplasm Proteins; Neoplastic Cells, Circulating; Spinal Neoplasms; Spine; Tissue Plasminogen Activator; Veins

The occurrence and distribution of components of coagulation pathways in situ were determined using immunohistochemical techniques applied to 10 cases of primary carcinoma of the breast, normal breast tissue obtained from two patients undergoing reductive mammoplasty, and three patients with benign breast tumors. Tumor cells stained for factor X and thrombomodulin but not for tissue factor, factor V, factor VII, or factor XIII. Rare nonneoplastic duct epithelial cells stained for thrombomodulin, but these tissues did not otherwise stain for any of these antigens. Macrophages within the tumor stroma stained for tissue factor, factor VII, and factor XIII but not for factor V or factor X. These features of macrophages were the same in malignant and nonmalignant breast tissue. Fibrinogen was present in abundance throughout the connective tissue in breast cancer but not in nonmalignant tissues. By contrast, no staining was observed using fibrin-specific antibodies. These results suggest that an intact coagulation pathway does not exist in breast cancer tissue and that thrombin capable of transforming fibrinogen to fibrin is not generated in significant amounts in this tumor type. While fibrin is not a feature of the connective tissue stroma in breast cancer, it is conceivable that the abundant fibrinogen present in the tumor connective tissue (and factor XIII present in connective tissue macrophages) might contribute to the structural integrity of breast tumor tissues.

Topics: Breast Neoplasms; Carcinoma; Carcinoma, Intraductal, Noninfiltrating; Factor VII; Factor X; Fibrin; Fluorescent Antibody Technique; Humans; Immunoenzyme Techniques; In Vitro Techniques; Thrombin; Transglutaminases

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

The infiltration of macrophages both within and at the margin of malignant neoplasms can be extensive, but their functions are not well defined. Definitions of the antigenic phenotype defined by various monoclonal antibodies may allow insight into macrophage function. Single and double immunoenzymatic labelling techniques were used to characterize sub-populations of macrophages both within and at the margin of breast carcinoma and colorectal carcinoma using a panel of antibodies. Factor XIII, previously identified in macrophage cytoplasm, was localized at the same sites. Two major groups of tumour-associated macrophages were identified; class II MHC+, CD11c+ macrophages predominated within the neoplasm, whereas CD14+ macrophages were the major population at the invasive margin. Factor XIII+ macrophages were also seen predominantly at the invasive margin. Phenotypic variation between macrophage sub-populations may reflect functional variation such that macrophages may be beneficial or detrimental for neoplastic growth. Factor XIII derived from macrophages may be important in stabilization of fibrin deposits associated with the neoplasm.

Topics: Antibodies, Monoclonal; Antigens, Differentiation; Breast Neoplasms; CD11 Antigens; Colorectal Neoplasms; Factor XIII; Female; Fibrin; Histocompatibility Antigens Class II; Humans; Immunohistochemistry; Macrophages; Phenotype

Fibrin formation and fibrinolysis were estimated in 89 breast cancer patients by measurement in plasma of Fibrin Fragment B beta 15-42 and Fibrinopeptide A (FPA), serum Fibrin(ogen) Degradation Products (FDPs) and plasminogen activator by Fibrin Plate Lysis Assay. Results were compared with (a) 26 patients with benign breast diseases; and (b) 45 healthy factory workers. FPA, FDP and B beta 15-42 levels were elevated in both breast cancer patients and benign disease patients, but there were no significant differences between these two groups. Cancer stage, patient age and smoking habits did not affect these results, but Oestrogen Receptor (ER) positive patients had higher B beta 15-42 values than ER negative patients (p = 0.017). These results show that fibrin formation is enhanced preoperatively in patients with either benign or malignant breast disease. The fibrinolytic response to activated coagulation may be relatively deficient in breast cancer. The roles of malignancy, stress and other factors in the causation of these abnormalities require further assessment.

Topics: Breast Neoplasms; Female; Fibrin; Fibrin Fibrinogen Degradation Products; Fibrinogen; Fibrinolysis; Fibrinopeptide A; Fibrinopeptide B; Humans; Peptide Fragments; Plasminogen Activators

In order to evaluate the efficacy of an anticoagulant treatment in neoplasia, we have looked for the existence and the possible role of hemostatic unbalance in patients affected by limited and uncomplicated thyroid and breast cancer by examining hemostasis in 25 patients. Our data allowed us to evidentiate an accelerated and increased fibrinoformation associated with the presence of plasminogen's activation inhibitor which overlaps the increased plasminogen's activators. We evidentiated also an increase of platelet functions in vitro and of their activation in vivo. These findings support the hypothesis of a possible role played by hemostasis alterations in the pathogenesis of thromboembolic complications, cancer growth and/or metastatization, and justify prophylactic and therapeutic anticoagulation.

Topics: Adult; Age Factors; Aged; Anticoagulants; beta-Thromboglobulin; Breast Neoplasms; Female; Fibrin; Fibrinolysis; Hemostasis; Humans; Male; Middle Aged; Platelet Aggregation; Thromboembolism; Thyroid Neoplasms

This paper reports studies on the interaction between human platelets, the plasma coagulation system, and two human tumor cell lines grown in tissue culture: Melanoma and breast adenocarcinoma. The interaction was monitored through the use of 125I-labelled fibrinogen, which measures both thrombin activity generated by cell-plasma interaction and fibrin/fibrinogen binding to platelets and tumor cells. Each tumor cell line activates both the platelets and the coagulation system simultaneously resulting in the generation of thrombin or thrombin-like activity. The melanoma cells activate the coagulation system through "the extrinsic pathway" with a tissue factor-like effect on factor VII, but the breast tumor seems to activate factor X directly. Both tumor cell lines activate platelets to "make available" a platelet-derived procoagulant material necessary for the conversion of prothrombin to thrombin. The tumor-derived procoagulant activity and the platelet aggregating potential of cells do not seem to be inter-related, and they are not specific to malignant cells.

Topics: Adenocarcinoma; Blood Platelets; Breast Neoplasms; Cell Line; Cells, Cultured; Factor VII; Factor X; Fibrin; Fibrinogen; Hemostasis; Humans; In Vitro Techniques; Iodine Radioisotopes; Melanoma

Study of 14 human infiltrating breast carcinomas revealed new features that shed light on the pathogenesis of tumor stroma formation and on host immunologic defense mechanisms. Fibrin deposits were observed in the stroma of all tumors, particularly at their growing edge. Fibrin may have contributed both to tumor angiogenesis and, with organization, to the formation of the fibrous stroma characteristic of these and other scirrhous carcinomas. We previously proposed similar mechanisms for several animal tumors. All breast carcinomas studied elicited some degree of lymphocytic response at the tumor periphery; lymphocytes penetrated the fibrous tumor stroma poorly, did not exit in significant numbers from central tumor vessels, and, even when greatly outnumbering tumor cells locally, appeared relatively ineffective at tumor cell killing. Microvascular endothelial cell damage was frequently observed and may have been responsible for zones of tumor infarction. Similar observations have been made in skin allografts and animal tumors where rejection was effected principally by microvascular damage and subsequent tissue infarction, not by lymphocyte contact with individual epithelial target cells.

Topics: Adenocarcinoma, Scirrhous; Adult; Aged; Animals; Breast Neoplasms; Carcinoma, Intraductal, Noninfiltrating; Female; Fibrin; Fluorescent Antibody Technique; Guinea Pigs; Humans; Lymphocytes; Microscopy, Electron; Middle Aged; Venules

Human mammary carcinoma cell cultures proliferated from primary explants in Eagle's essential medium (MEM) supplemented with insulin, fetal calf serum (FCS) and/or human alpha-a1-antitrypsin. Human mammary carcinoma cells differed from normal mammary epithelial cells by the following catalytic activities: a. Thymidine uptake into the carcinoma cells was 6 to 10 fold greater, whereas thymidine conversion to CO2 was half to one fifth that of normal cells. b. The nucleolytic activity patterns of the mammary carcinoma cells preferred polycytydylic acid and double helical polynucleotides, whereas those of the normal mammary cells preferred polyuridylic acid and had no effect on double helical polynucleotides. c. The polymerase activity most evident in mammary carcinoma cells is a hybrid-dependent DNA polymerase which is guided by the ribo-strand of the template poly (rA) . poly(dT). In contrast the all-ribo template poly (rA) . poly(rU) showed little activity. d. There was slight or statistically non-significant difference between the amino acid composition of material cleaved from mammary carcinoma cells prepared from tumor tissues and from cells cultivated 10 months in vitro. e. There was no difference between the molar proportions of the carbohydrate components of the cell membrane from fresh tumor tissue and long term in vitro cultivated cells. f. The granules from long term in vitro cultured mammary carcinoma cells contained high collagenolytic, caseinolytic, fibrinolytic and esterolytic activities.

Topics: Amino Acids; Breast Neoplasms; Carbohydrate Metabolism; Cell Nucleus; Cells, Cultured; Culture Media; DNA-Directed DNA Polymerase; Enzyme Activation; Female; Fibrin; Humans; In Vitro Techniques; Thymidine; Time Factors

Topics: Breast Neoplasms; Colonic Neoplasms; Esophageal Neoplasms; Fibrin; Fibrinogen; Fibrinolysis; Gallbladder Neoplasms; Humans; Lymphatic Metastasis; Neoplasms; Pancreatic Neoplasms; Rectal Neoplasms; Stomach Neoplasms

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: Anticoagulants; Breast Neoplasms; Female; Fibrin; Humans; Neoplasm Metastasis; Venoms

Topics: Adenofibroma; Basement Membrane; Breast; Breast Neoplasms; Carcinoma; Carcinoma, Intraductal, Noninfiltrating; Connective Tissue; Epithelium; Female; Fibrin; Fibroblasts; Humans; Microscopy, Electron

Topics: Adenocarcinoma; Adenoma; Adrenal Gland Neoplasms; Breast Diseases; Breast Neoplasms; Female; Fibrin; Fluorescent Antibody Technique; Genital Neoplasms, Female; Hodgkin Disease; Humans; Lymph Nodes; Lymphatic Diseases; Melanoma; Methods; Neoplasms; Neoplasms, Nerve Tissue; Ovarian Neoplasms; Sarcoma; Stomach Neoplasms; Thyroid Neoplasms; Tuberculosis

Topics: Aged; Ascites; Breast; Breast Neoplasms; Cell Division; Culture Techniques; Epithelium; Female; Fibrin; Humans; Middle Aged; Neoplasms; Nylons; Ovarian Neoplasms; Pleura; Rectal Neoplasms