Human colon carcinoma cells synthesize and secrete alpha 1-proteinase inhibitor.
Our previous results have shown that tumor cell-secreted procathepsin B can be activated by neutrophil elastase in vitro. In the present study, we addressed two questions: 1. Can neutrophil elastase be detected in human colon carcinomas, and 2. Does the co-culture of human colon carcinoma cells with neutrophils generate a cathepsin B-dependent pericellular proteolysis as assessed with radiolabeled laminin? We show that neutrophil elastase is present in colon carcinoma tissue and that its level is in good agreement with the degree of tissue infiltration by neutrophils. In co-culture experiments, elastase is released by neutrophils in a cell number dependent way, but no activation of tumor cell-secreted procathepsin B could be observed. In addition, the degradation of radiolabeled laminin by neutrophil proteinases was markedly decreased in the presence of tumor cells. These findings prompted us to search for a tumor cell-secreted elastase inhibitor. We show by enzyme activity measurements, gelatin-zymography, immunoblotting and RT-PCR that colon carcinoma cells synthesize and secrete alpha 1-proteinase inhibitor, a functional inhibitor of neutrophil elastase. The importance of this finding in the context of pericellular activation of tumor cell-secreted procathepsin B by neutrophil elastase is discussed.
Keppler
D
D
Experimental Pathology Unit, Swiss Institute for Experimental Cancer Research (ISREC), Epalinges, Switzerland.
Markert
M
M
Carnal
B
B
Berdoz
J
J
Bamat
J
J
Sordat
B
B
D015152:Blotting, Northern; D015139:Blotting, Southern; D002277:Carcinoma / Q000201:enzymology*; D002401:Cathepsin B / Q000378:metabolism; D018920:Coculture Techniques; D003110:Colonic Neoplasms / Q000201:enzymology*; D017077:Culture Media, Conditioned; D004789:Enzyme Activation / Q000502:physiology; D005109:Extracellular Matrix / Q000201:enzymology; D006801:Humans; D007797:Laminin / Q000378:metabolism; D019272:Leukocyte Elastase / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology / Q000378:metabolism; D010196:Pancreatic Elastase / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D000515:alpha 1-Antitrypsin / Q000096:biosynthesis* / Q000378:metabolism
D017077:Culture Media, Conditioned; D007797:Laminin; D000515:alpha 1-Antitrypsin; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase; D002401:Cathepsin B