Immunolocalization of collagenase in neoplastic epithelial cells.
The distribution of interstitial collagenase in a rat mammary carcinoma model system has been studied by immunocytochemistry. Rabbit antibodies were raised against collagenase from neoplastic epithelial cells which were derived from an anaplastic, invasive, rat mammary carcinoma (BC1). Specificity of the antibodies was determined by Western blot analysis which showed reactivity with the inactive procollagenase from conditioned culture medium of BC1 cells as well as with purified, active BC1 collagenase. Anti-BC1 collagenase antibodies did not recognize BC1 collagenase entrapped by the inhibitor, rat alpha-2-macroglobulin (alpha 2M), or collagenase derived from TPA-stimulated human fibroblasts. Anti-human fibroblast collagenase antibodies did not recognize BC1 collagenase, suggesting that the human-mesenchymal and rat-epithelial enzymes are immunologically distinct molecules. Collagenase was immunolocalized intracellularly in BC1 cells cultured in the presence of monensin. Neither BC1 collagenase, alpha 2M nor enzyme-inhibitor complexes were demonstrated in or around invading tumours by immunostaining of tissue sections of rat mammary carcinomas.
Whitelock
J M
JM
Department of Cellular Pathology, University of Technology, Sydney, Gore Hill, NSW, Australia.
O'Grady
R L
RL
Lyons
J G
JG
D000818:Animals; D000912:Antibodies, Neoplasm / Q000096:biosynthesis / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D002277:Carcinoma / Q000032:analysis / Q000201:enzymology*; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000032:analysis / Q000201:enzymology; D005260:Female; D007124:Immunoenzyme Techniques; D008325:Mammary Neoplasms, Experimental / Q000032:analysis / Q000201:enzymology*; D003012:Microbial Collagenase / Q000032:analysis* / Q000378:metabolism; D008985:Monensin / Q000494:pharmacology; D009368:Neoplasm Transplantation; D011817:Rabbits; D051381:Rats; D014407:Tumor Cells, Cultured; D000511:alpha-Macroglobulins / Q000378:metabolism
D000912:Antibodies, Neoplasm; D000511:alpha-Macroglobulins; D008985:Monensin; D003012:Microbial Collagenase