carboxypeptidase-b and Adenocarcinoma

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

To clarify the difference in the protein composition of pancreatic juice between patients with pancreatic cancer and normal controls, the proteins of pure pancreatic juice from three cases of cancer of the head of the pancreas and six apparently healthy adults were analyzed by two-dimensional gel electrophoresis (two-DE) followed by silver staining. Two minor proteins of Mr 59,000 and Mr 78,000 present in all the three patients were not detected in the six controls. We have identified the minor proteins with Mr 59,000 and Mr 78,000 as alpha 1-antitrypsin and transferrin, respectively, using Western blotting with anti-alpha 1-antitrypsin and anti-transferrin antibodies. In addition, serum albumin also identified by antibody binding was abundantly present in patients compared to controls. The increase in the amount of serum albumin in the patient was also confirmed by a quantitative analysis using SDS-PAGE and gel densitometry. The data indicate that not only serum albumin but also alpha 1-antitrypsin and transferrin are increased in the pancreatic juice of the patients with pancreatic cancer, as compared with apparently healthy adults. The data also suggests that the analysis of pancreatic juice proteins by two-DE with silver staining is useful for the diagnosis of pancreatic diseases.

Topics: Adenocarcinoma; alpha 1-Antitrypsin; Amylases; Carboxypeptidase B; Carboxypeptidases; Carboxypeptidases A; Densitometry; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Enzyme Precursors; Humans; Lipase; Pancreatic Juice; Pancreatic Neoplasms; Proteins; Serum Albumin; Sodium Dodecyl Sulfate; Staining and Labeling; Transferrin