thromboplastin and Carcinoma--Squamous-Cell

Tumor-specific biomarkers are a prerequisite for the development of targeted imaging and therapy in oral squamous cell carcinoma (OSCC). urokinase-type Plasminogen Activator Receptor (uPAR), Tissue Factor (TF) and Epidermal Growth Factor Receptor (EGFR) are three biomarkers that exhibit enhanced expression in many types of cancers, and have been investigated as potential biomarkers for targeted strategies and prognostication. The aim of the study was to investigate the expression patterns of uPAR, TF and EGFR and their potential prognostic value in OSCC.. Immunohistochemical expression of uPAR, TF and EGFR in tumor resection specimens from 191 patients with primary OSCC was analyzed. Overall (OS) and disease-free survival (DFS) was calculated. Associations between biomarker expression, clinicopathological factors and patient survival was analyzed using the Cox proportional hazards model for univariate and multivariate analysis, log rank and Kaplan-Meier statistics.. uPAR and TF exhibited a highly tumor-specific expression pattern while EGFR also showed expression in normal tissues outside the tumor compartment. The overall positive expression rate of uPAR, TF and EGFR was 95%, 58% and 98%, respectively. High uPAR expression across the entire cohort was negatively associated with OS (p = 0.031, HR = 1.595 (95%CI 1.044-2.439)) in univariate analysis. The 5-year OS for high and low uPAR expression was 39% and 56%, respectively. The expression of TF and EGFR was not associated with survival outcome.. This study may suggest that uPAR and TF could potentially be attractive targets for molecular imaging and therapy in OSCC due to high positive expression rates and tumor-specific expression patterns. High uPAR expression was significantly associated with a reduced survival. uPAR seems to be a prognostic biomarker in oral cancer.

Topics: Adult; Aged; Aged, 80 and over; Apoenzymes; Biomarkers, Tumor; Carcinoma, Squamous Cell; Disease-Free Survival; ErbB Receptors; Female; Gene Expression Regulation, Neoplastic; Humans; Male; Middle Aged; Mouth Neoplasms; Proportional Hazards Models; Receptors, Urokinase Plasminogen Activator; Thromboplastin; Young Adult

: Microvesicles associated with tissue factor (TF) may play a role in cancer-related venous thromboembolism; however, not much is known about their interaction with the tumour stroma, especially the endothelium or any procoagulant changes seen because of this interaction. Using a head and neck squamous cell carcinoma line (UMSCC81B) and human umbilical vein endothelial cells (HUVECs), this study explored the interaction of cancer microvesicles released into cell culture media with endothelial cells in vitro, and assessed the procoagulant activity resulting from this interaction. Cell-free media containing UMSCC81B cancer microvesicles supported coagulation in a concentration-dependent manner, suggesting TF and microvesicle presence, this media was then added to HUVECs and flow cytometry analysis showed a subpopulation of HUVECs that had acquired a significantly high expression of TF, which was dependent upon the concentration of UMSCC81B media containing microvesicles present and confocal microscopy confirmed HUVECs associated with labelled microvesicles. The range of TF-positive HUVECs was determined to be 0, 4.2(±1.4), 12.5(±3.72), and 45.9(±18.7)% for microvesicle-positive media concentration of 0, 25, 50, and 100%, respectively, which resulted in decreasing prothrombin values of more than 600 (no clot), 126.4, 65.8, and 47.8 s. Our results demonstrate that procoagulant microvesicles shed by UMSCC81B induced a procoagulant effect in HUVECs through increased clotting activity and cell membrane surface expression of TF.

Topics: Blood Coagulation; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell-Derived Microparticles; Endothelial Cells; Endothelium, Vascular; Head and Neck Neoplasms; Human Umbilical Vein Endothelial Cells; Humans; Squamous Cell Carcinoma of Head and Neck; Thromboplastin; Venous Thromboembolism

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

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

Neoadjuvant chemotherapy (NACT) improves the prognosis of patients with esophageal cancer who respond, but it is not effective in nonresponders. Therefore, it is crucial to establish a reliable method of predicting response before initiation of chemotherapy. Hypercoagulability, which is thought to be because of upregulation of tissue factor (TF) in cancer cells, was reported to be associated with chemoresistance. The aim of this study was to investigate the association between TF expression and response to NACT in esophageal cancer.. In 67 patients with advanced esophageal cancer, TF expression in pretreatment biopsy samples was evaluated immunohistochemically and correlated with clinicopathologic factors and response to chemotherapy.. TF was expressed by 43.3% of the tumors, but there were no correlations observed with any clinicopathologic parameters examined. Clinical and histologic responses to chemotherapy were significantly worse in TF-positive patients compared with TF-negative patients. Multivariate analysis revealed that TF expression was significantly associated with a poor clinical response (P = 0.0431). TF expression was also independently associated with poor progression-free survival (P = 0.0353).. TF expression levels in pretreatment biopsy samples are useful for predicting response to NACT in advanced esophageal cancer. Further studies of mechanisms underlying the relationship between TF expression and chemosensitivity are needed.

Topics: Aged; Biopsy; Carcinoma, Squamous Cell; Disease Progression; Disease-Free Survival; Drug Resistance, Neoplasm; Esophageal Neoplasms; Esophagus; Female; Humans; Male; Middle Aged; Neoadjuvant Therapy; Neoplasm Grading; Predictive Value of Tests; Prognosis; Thrombophilia; Thromboplastin

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

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

It has been demonstrated that tissue factor (TF) may be involved in the tumor-derived procoagulatory status and angiogenic modulation in certain solid tumors. In the present study, we examined immunohistochemical localisation of TF in esophageal squamous cell carcinomas (ESCC) from 103 patients. TF immunopositivity was found in 91.3% of all tumor sections, while normal esophageal tissues were immunonegative. Patients were divided into a low TF immunoreactivity group (9 cases of negative and 48 cases of weak positive) and a high TF immunoreactivity group (35 cases of moderate positive and 11 cases of strong positive). TF immunoreactivity was significantly correlated to the presence of distant metastasis (P=0.0014), while it was not correlated to patient's gender, age, tumor size, depth of tumor invasion or lymph node metastasis. Survival analysis revealed that the overall survival rate in the patients that had high TF immunoreactivity was significantly poorer than those with low TF immunoreactivity (P=0.0094). Univariate analysis demonstrated that tumor size (P=0.0095), depth of tumor invasion (P=0.0050), lymph node metastasis (P=0.0045) and distant metastasis (P<0.0001) were effective predictors of prognosis in patients. However, only distant metastasis could independently predict patients' outcomes by the analysis of multivariate proportional hazards regression (P=0.0043). Furthermore, the intratumoral microvessel density (MVD), evaluated by CD34 immunolabeling, indicated that MVD was positively correlated to the TF immunoreactivity (P=0.0056). It is concluded that TF immunopositivity in ESCC tissues is strongly correlated to the intratumoral angiogenesis and to poor patient prognosis.

Topics: Adult; Age Factors; Aged; Antigens, CD34; Carcinoma, Squamous Cell; Esophageal Neoplasms; Female; Hemostatics; Humans; Lymphatic Metastasis; Male; Microvessels; Middle Aged; Multivariate Analysis; Neovascularization, Pathologic; Proportional Hazards Models; Sex Factors; Survival Analysis; Thromboplastin

ErbB oncogenes drive the progression of several human cancers. Our study shows that in human carcinoma (A431) and glioma (U373) cells, the oncogenic forms of epidermal growth factor receptor (EGFR; including EGFRvIII) trigger the up-regulation of tissue factor (TF), the transmembrane protein responsible for initiating blood coagulation and signaling through interaction with coagulation factor VIIa. We show that A431 cancer cells in culture exhibit a uniform TF expression profile; however, these same cells in vivo exhibit a heterogeneous TF expression and show signs of E-cadherin inactivation, which is coupled with multilineage (epithelial and mesenchymal) differentiation. Blockade of E-cadherin in vitro, leads to the acquisition of spindle morphology and de novo expression of vimentin, features consistent with epithelial-to-mesenchymal transition. These changes were associated with an increase in EGFR-dependent TF expression, and with enhanced stimulation of vascular endothelial growth factor production, particularly following cancer cell treatment with coagulation factor VIIa. In vivo, cells undergoing epithelial-to-mesenchymal transition exhibited an increased metastatic potential. Furthermore, injections of the TF-blocking antibody (CNTO 859) delayed the initiation of A431 tumors in immunodeficient mice, and reduced tumor growth, vascularization, and vascular endothelial growth factor expression. Collectively, our data suggest that TF is regulated by both oncogenic and differentiation pathways, and that it functions in tumor initiation, tumor growth, angiogenesis, and metastasis. Thus, TF could serve as a therapeutic target in EGFR-dependent malignancies.

Topics: Animals; Cadherins; Carcinoma, Squamous Cell; Cell Differentiation; Cell Line, Tumor; Epithelial Cells; ErbB Receptors; Flow Cytometry; Glioma; Humans; Mesoderm; Mice; Mice, SCID; Neoplasm Metastasis; Neovascularization, Pathologic; Thromboplastin; Up-Regulation; Vascular Endothelial Growth Factor A; Vimentin

Topics: AC133 Antigen; Antigens, CD; Caco-2 Cells; Carcinoma, Squamous Cell; Cell Line, Tumor; Glycoproteins; Humans; Neoplastic Stem Cells; Peptides; Thromboplastin

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

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

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

It has been established that the only source of tissue thromboplastin activity in homogenate of cells (HeLa, L-41, RAMT, Hep-2 and FL) grown in synthetic nutrient media are permolecular formations represented by fragments of cellular membranes Thromboplastic activity of nutrient media is also associated with these formations, as fragments of cellular membranes enter the media during cell cultivation. The removal of all permolecular formations from homogenate of cells, from the medium in which they are cultivated, allows determination of the topography of tissue coagulation factors.

Topics: Amnion; Animals; Callitrichinae; Carcinoma, Squamous Cell; Cattle; Cell Line; Culture Media; Female; HeLa Cells; Humans; In Vitro Techniques; Kidney; Laryngeal Neoplasms; Leukemia; Male; Thromboplastin

Generation of thromboplastin by monocytes has been shown to play a vital role in hypercoagulable states seen in malignancy. The purpose of this study was to compare the procoagulant activity in cancer patients and controls. Recalcification times (RT) of whole blood from 19 normal volunteers, 8 patients with benign polyps, 12 patients previously treated by surgery for head and neck (H&N) or colon cancer, and 13 untreated patients with various stages of H&N or colon cancer were determined. Tests were performed with and without stimulation with Escherichia coli endotoxin. The mean RT in saline (RTS) of untreated patients with early cancer (4.58 +/- 0.83 min) and that of patients with advanced cancer (5.23 +/- 1.16 min) were lower than that of controls (6.55 +/- 0.82 min), P less than 0.01 and P less than 0.05, respectively. The RTS of patients previously treated and of those with benign polyps were no different from those of controls. Activation with endotoxin significantly lowered the recalcification times (RTE) in the early (3.90 +/- 0.58 min) and advanced cancer patients (4.23 +/- 0.66 min) compared to the RTE of controls (5.69 +/- 0.75 min, P less than 0.01 for both groups) as well as compared to those with benign tumors, P less than 0.05. The mean RTE of previously treated patients (4.72 +/- 0.58 min) was also lower than that of controls, P less than 0.05. Our results suggest that RT is significantly reduced in cancer patients compared to that of controls. Furthermore, monocyte activation with endotoxin may enable us to distinguish cancer patients from controls as well as from those with benign tumors.

Topics: Adenoma; Adult; Blood Coagulation Tests; Carcinoma, Squamous Cell; Colonic Neoplasms; Head and Neck Neoplasms; Humans; Male; Middle Aged; Monocytes; Neoplasms; Thromboplastin

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

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

Thromboplastic and fibrinolytic activities of V2 and V7 carcinomas, the two transplantable rabbit tumors of the same viral origin, were studied in relation to fibrin deposition and thrombus formation in the tumors. Thromboplastic activity of V7 carcinoma was comparatively high, while that of V2 carcinoma was as low as that of muscle tissue. More fibrin deposits in the stroma and more thrombi in the small vessels were found at the advancing border of V7 carcinoma than that of V2 carcinoma. These differences might be associated with higher thromboplastic activity of V7 carcinoma than that of V2 carcinoma. Fibrinolytic activity of both tumors was high and it was confirmed to be localized in the tumor cells by Todd's method. Fibrin deposits in the stroma were found more abundantly somewhat apart from the advancing border of the tumor nests of both tumors. It was suggested that plasmin activated by plasminogen activator released locally from the tumor cells might digest fibrin deposited in the stroma just close to the tumor nests.

Topics: Animals; Carcinoma, Squamous Cell; Fibrin; Fibrinolysis; Male; Neoplasms, Experimental; Plasminogen Activators; Rabbits; Thromboplastin; Thrombosis

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

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