fibrin and Cystadenocarcinoma

The essential role of urokinase-type plasminogen activator (uPA) in tumor invasion and metastasis stresses the necessity of a fine-tuned cellular control over its expression. It has been shown that changes in uPA directly correlate with changes in cell invasiveness. We examined the role of Rel-related proteins in uPA synthesis by human ovarian cancer cells by inhibiting their expression using the antisense (AS) oligodeoxynucleotide (ODN) technology. Exposure of OV-MZ-6 cells to 10 microM phosphorothioate (PS)-derivatized AS-ODN directed to Rel A led to a maximal 50% decrease of uPA antigen in cell lysates and a 70% reduction in cell cultures supernatants accompanied by a significant transient decline in uPA mRNA levels. Antisense-PS-ODN directed to NF-kappa B1 (p50) or c-rel had no effect on uPA protein expression. AS-PS-ODN directed to Rel A also affected the proteolytic capacity of OV-MZ-6 cells reflected by an approximately 70% decrease in the fibrinolytic capacity of the cells within 24 h compared to untreated controls. AS-PS-ODN directed to I kappa B alpha expression increased uPA in cell culture supernatants up to 50%. uPA receptor (uPAR) production and synthesis of plasminogen activator inhibitor type-1 (PAI-1) were not altered by either AS-PS-ODN applied. Western blot and gel retardation analyses revealed constitutive expression of Rel-related proteins in nuclear protein extracts of OV-MZ-6 cells. Thus these proteins seem to be implicated in uPA regulation and may thereby contribute to tumor spread and metastasis.

Topics: Base Sequence; Cystadenocarcinoma; Female; Fibrin; Flow Cytometry; Gene Expression; Gene Transfer Techniques; Humans; Molecular Sequence Data; NF-kappa B; Oligonucleotides, Antisense; Ovarian Neoplasms; Plasminogen Activator Inhibitor 1; Polymerase Chain Reaction; RNA-Directed DNA Polymerase; RNA, Messenger; Transcription Factor RelA; Tumor Cells, Cultured; Urokinase-Type Plasminogen Activator

The aim of this study was to evaluate the type of enzymes involved in tumor-associated fibrinolysis of the stroma component fibrin in ovarian cancer patients. For this purpose, the high-molecular-mass fibrin degradation products (HMM-XDP) were isolated from malignant ascitic fluid by protamine sulfate precipitation and further purified by gel filtration and acid precipitation. After reduction with 2-mercaptoethanol, the peptide chain components were separated by reverse-phase high-performance liquid chromatography (RP-HPLC). The nature of these components was elucidated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid sequence analysis and compared with fibrin-derived fragments formed in vitro. The results indicate that plasmin is the essential protease involved in the degradation of the stroma-derived fibrin portion found in ovarian cancer ascites.

Topics: Amino Acid Sequence; Ascitic Fluid; Chromatography, High Pressure Liquid; Cystadenocarcinoma; Electrophoresis, Polyacrylamide Gel; Factor XIII; Female; Fibrin; Fibrinogen; Fibrinolysin; Humans; Molecular Sequence Data; Ovarian Neoplasms; Peptide Hydrolases; Thrombin

Covalently linked heterogeneous fibrin-fibronectin compounds were detected in ascitic fluid of 31 patients with advanced ovarian cystadenocarcinoma by means of enzyme-linked immunosorbent assay techniques, immunoaffinity chromatography, and Western blot analysis. Deposition of fibrin and fibronectin could also be demonstrated immunohistochemically in Carnoy-fixed tissue sections. Fibrin and fibronectin were found in the tumor stroma within tumor nests and more prominently in stroma surrounding the tumor nests. The association of fibrin and fibronectin was especially pronounced in the stroma surrounding the tumor islands. Fibronectin was also found to be associated with stroma cells. Areas within the tumor stroma showed superimposed staining for both fibrin and fibronectin supporting the assumption that the covalently linked fibrin-fibronectin conjugates found in ascitic fluid may stem from the provisional tumor stroma by proteolytic release.

Topics: Ascitic Fluid; Cystadenocarcinoma; Enzyme-Linked Immunosorbent Assay; Female; Fibrin; Fibronectins; Humans; Immunohistochemistry; Molecular Weight; Ovarian Neoplasms; Plasminogen Activators; Urokinase-Type Plasminogen Activator

Fibrin degradation products (F.D.P.) were determined in the serum of 163 women in whom ovarian tumours had been suspected on palpation at gynaecological examination and who were afterwards examined by laparoscopy or subjected to laparotomy. F.D.P. were found in the serum (0.5-30 mg/100 ml) of 23 (72%) out of 32 patients with malignant tumours. Of 131 patients with benign findings F.D.P. (traces to 2 mg/100 ml) were found in six (4.5%), and in most of these the occurrence of F.D.P. could be explained on other clinical grounds. The findings suggest that the examination of F.D.P. in suspected malignant ovarian tumour may be of diagnostic value.Determination of F.D.P. in malignant ascitic fluid showed very high values, ranging between 40 and 350 mg/ 100 ml. This argues for the occurrence of F.D.P. in the blood being due to an extravascular breakdown of fibrin caused by tumour cells, but they may also be due to thromboplastic and fibrinolytic agents from the tumour entering the blood stream.

Topics: Ascitic Fluid; Carcinoma; Cystadenocarcinoma; Cystadenoma; Electrophoresis; Female; Fibrin; Granulosa Cell Tumor; Humans; Laparoscopy; Laparotomy; Liposarcoma; Mesonephroma; Molecular Weight; Ovarian Neoplasms