title: Degradation of intact basement membranes by human and murine tumor enzymes.
abstract: Homogenates from malignant tumors, obtained from surgery specimens or from transplants of Walker 256 carcinosarcoma in rats, contained an enzyme activity capable of degrading intact 3H-acetylated basement membranes from bovine lens. The enzyme activity from murine tumor was purified about 7500-fold by (NH4)2SO4 fractionation, ion exchange and gel chromatography. The apparent molecular weight of the purified enzyme was approximately 50,000. The rate of degradation of 3H-labelled basement membrane by the murine tumor enzyme was reduced by addition of excess type IV collagen, but not of excess type I, type III or type V collagen. These results suggested specificity of this enzyme for type IV collagen. Inhibitors of serine proteinases, thiol proteinases and soybean trypsin inhibitor were without effect on the enzyme activity. Chelators such as 1,10-phenanthroline or EDTA reduced the activity to control levels, indicating that the enzyme activity was due to a metalloproteinase. Chromatographic and electrophoretic separation of the enzymatic products from 3H-labelled basement membrane and type IV collagen indicated that the enzyme activity was due to a type IV collagenase. The use of basement membrane in the native physiological state as a substrate for the study of basement membrane-degrading activity by homogenates of solid malignant tumors offers an in vitro model for the investigation of the metastatic potential of these tumors.
authors.affiliation: Department of Pharmacology, University of Patras Medical School, Greece.
authors.forename: G
authors.initials: G
authors.lastname: Karakiulakis
authors.forename: E
authors.initials: E
authors.lastname: Missirlis
authors.forename: A
authors.initials: A
authors.lastname: Aletras
authors.forename: M E
authors.initials: ME
authors.lastname: Maragoudakis
mesh_terms: D000107:Acetylation; D000818:Animals; D001485:Basement Membrane / Q000378:metabolism*; D002279:Carcinoma 256, Walker / Q000201:enzymology*; D002417:Cattle; D002460:Cell Line; D006801:Humans; D051379:Mice; D008970:Molecular Weight; D051381:Rats; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured / Q000201:enzymology*