OBJECTIVETo evaluate the effect of tissue factor (TF) in extravascular migration of fibrosarcoma cells and hematogenous metastasis.

METHODSThe expression of tissue factor in fibrosarcoma HT1080 cells was analyzed by flow cytometry. The extravascular migration of fibrosarcoma cells was observed in a constructed monolayer vascular endothelial cells and extra-cellular matrix model.

RESULTTissue factor was highly expressed in HT1080 cells. HT1080 migrated and passed through the monolayer vascular endothelial cells to the collagen gel in a time-dependent manner. Anti-TF antibody inhibited extravascular migration of fibrosarcoma cells and the inhibition was concentration-dependent (P<0.05).

CONCLUSIONTissue factor may enhance hematogenous metastasis through extravascular migration of fibrosarcoma cells.

Cell Movement ; Cells, Cultured ; Endothelial Cells ; Fibrosarcoma ; metabolism ; pathology ; Humans ; Neoplasm Metastasis ; Thromboplastin ; metabolism ; physiology

Acute Coronary Syndrome ; blood ; Aged ; Cell Membrane ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Thromboplastin ; metabolism

Blood Coagulation ; physiology ; Blood Platelets ; metabolism ; Cell Communication ; Factor X ; metabolism ; Humans ; Leukocytes ; metabolism ; Thromboplastin ; metabolism ; physiology

Malignant tumor is one of the important acquired risk factors of venous thromboembolism (VTE). As the transmembrane receptor of coagulation factor Ⅶ and activated coagulation factor Ⅶa
Humans ; Neoplasms/complications* ; Risk Factors ; Thromboplastin/metabolism* ; Thrombosis ; Venous Thromboembolism/physiopathology*

Adolescent ; Adult ; Child ; E-Selectin ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Thromboplastin ; metabolism ; Young Adult ; beta-Thalassemia ; blood

The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.
Alternative Splicing ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Thromboplastin ; genetics ; metabolism ; Tumor Cells, Cultured ; U937 Cells

OBJECTIVETo investigate the effect of simulated microgravity on the proliferation of human monocytic cells THP-1 and the expression of tissue factor (TF) mRNA.

METHODSTHP-1 cells were cultured under a simulated microgravity environment using the rotating cell culture system (RCCS). The changes in the cell proliferation after microgravity culture were assessed by cell counting and cell cycle analysis with flow cytometry. RT-PCR was used to detect the changes in the expression of TF mRNA in THP-1 cells.

RESULTSCulture under simulated microgravity resulted in a significant decrease in the cell number of THP-1 cells in comparison with that of the control cells (P<0.01). After a 24-h culture under microgravity, the G0-Gl phase cells increased from the control level of (46.57∓1.64)% to (67.64∓2.71)% (P<0.05). The cells in both groups showed a low level of TF mRNA expression in the absence of LPS stimulation. A 4-h stimulation with LPS caused up-regulated expression of TF mRNA in both cells, but the microgravity group showed a significantly smaller increase in the expression (2.301∓0.179) than the control group (9.210∓1.328) (P<0.05).

CONCLUSIONMicrogravity can inhibit the proliferation of THP-1 cells and suppress the cellular expression of TF mRNA.

Cell Proliferation ; Cells, Cultured ; Humans ; Monocytes ; cytology ; metabolism ; RNA, Messenger ; genetics ; Thromboplastin ; genetics ; metabolism ; Weightlessness ; Weightlessness Simulation

OBJECTIVETo explore whether normal platelet contains tissue factor (TF), and the significance of platelet-associated TF (PATF).

METHODSPlatelets were isolated by Sepharose 2B gel column. ELISA was used to detect the TF content in the lysates of washed platelets. Procoagulant activity of PATF was measured by one stage clotting time assay. The mRNA of TF was detected by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSA certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in the washed-platelet lysates. Upon activation by collagen, platelets released TF and caused a marked increase in TF level in plasma (P <0.05). Resting platelets had no TF procoagulant activity, while procoagulant activity of platelets activated by collagen increased significantly, which could be blocked by TF McAb and poor VII plasma. TF mRNA could not be detected in washed platelets. TF content in platelets from patients with coronary heart disease was significantly higher than that from normal controls (P < 0.05). Resting platelets from the patients showed a higher procoagulant activity, which could be inhibited by TF McAb.

CONCLUSIONPlatelets contain TF and the latter released by activated platelet was functionally active. Platelet itself might not synthesize TF. Protein content and procoagulant activity of PATF in patients with coronary heart disease were higher than that in controls. All these indicate that platelet may be involved in coagulation and thrombosis by releasing TF.

Adolescent ; Adult ; Blood Platelets ; chemistry ; Coronary Artery Disease ; metabolism ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Platelet Activation ; Thromboplastin ; metabolism ; physiology

This study was aimed to investigate the expression and activity of membrane surface tissue factor (TF) of monocytes and platelets in peripheral blood cells from patients with cerebral infarction and their clinical significance. The TF expressions in monocytes and platelets from 25 patients with cerebral infarction were detected by flow cytometry, the TF activity was detected by chromogenic reaction method, and compared with 24 normal people used as control. The results showed that the TF expressions of monocytes and platelets in peripheral blood cells from patients with cerebral infarction were significantly higher than that in normal controls (p<0.01), and TF activity was also higher in patients than that in controls (p<0.01). In conclusion, the expression and activity of membrane surface in patients with cerebral infarction were enhanced, the hematocyte-derived tissue factor as a trigger in coagulation pathway is involved in pathological thrombosis in patients with cerebral infarction.
Aged ; Blood Cells ; metabolism ; Case-Control Studies ; Cerebral Infarction ; blood ; metabolism ; Erythrocyte Membrane ; metabolism ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Monocytes ; metabolism ; Thromboplastin ; metabolism

This study was purposed to detect the expression level of human endothelial cell tissue factor (hECTF) and concentration of IL-2, and to investigate the alterations of hECTF and IL-2 after using immunosuppressive agent (cyclosporin, CsA) and explore the significance of endothelial cell (EC) lesion and abnormal expression of tissue factor (TF) in GVHD. Human endothelial cells and allogeneic lymphocytes were mixed and cultured as well as were cocultured with CsA for 4-6 hours in vitro, then the expression level of hECTF was detected by flow cytometry and RT-PCR, the concentration of IL-2 in supernatant was assayed by ELISA. The experiment was divided into 4 groups: 1st group--non-mixed-cultured group (negative control group), 2nd group - mixed-cultured group (positive control group), 3rd group - mixed-cocultured group with 1 µg/ml CsA and 4th group--mixed-cocultured group with 2 µg/ml CsA. The results showed that as compared with non-mixed-cultured group (negative control group), the expression level of hECTF and concentration of IL-2 in another 3 groups significantly increased (p<0.01), while as compared with positive control group, the expression level of hECTF and concentration of IL-2 in cocultured groups with CsA both decreased (p<0.01). It is concluded that the lesion of EC and abnormal expression of TF play a crucial role in GVHD, among which the high expression of TF after being stimulated by donor's lymphocytes may be the key step for occurrence and progression of GVHD.
Cells, Cultured ; Coculture Techniques ; Cyclosporine ; pharmacology ; Endothelial Cells ; metabolism ; Graft vs Host Disease ; metabolism ; pathology ; Humans ; Interleukin-2 ; metabolism ; Lymphocytes ; cytology ; Thromboplastin ; metabolism