Suppression of type I collagenase expression by antisense RNA in melanoma cells results in reduced synthesis of the urokinase-type plasminogen activator receptor.
Previously we reported that suppression of type I collagenase synthesis in human melanoma cells with antisense RNA significantly reduced proteolysis of type I and type IV collagen matrices (Durko et al., 1997, Biochim. Biophys. Acta 1356, 271). Because plasmin is a major activator of the type I collagenase, we assessed the impact of type I collagenase suppression on the urokinase/plasmin system of proteolysis. Gel zymography revealed the appearance of two new caseinolytic bands of Mr 81-83000 in conditioned media of type I collagenase-depleted, but not of wild-type cells and these were identified as plasmin bands. This increased extracellular plasmin activity coincided with reduced membrane-associated plasminogen levels and decreased expression of the urokinase-type plasminogen activator receptor at both the mRNA (up to 83% reduction) and cell-surface (up to 48% reduction) levels, while urokinase mRNA levels remained unchanged. The results indicate that in these cells the urokinase/plasmin system is regulated by type I collagenase levels.
Durko
M
M
Department of Surgery, McGill University and The Royal Victoria Hospital, Montreal, Quebec, H3A 1A1, Canada.
Brodt
P
P
D017364:Collagenases / Q000235:genetics*; D017077:Culture Media, Conditioned; D005341:Fibrinolysin / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D020781:Matrix Metalloproteinase 1; D008545:Melanoma / Q000235:genetics* / Q000378:metabolism*; D010960:Plasminogen Activators / Q000378:metabolism; D016372:RNA, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000235:genetics; D055293:Receptors, Urokinase Plasminogen Activator; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics / Q000378:metabolism
D017077:Culture Media, Conditioned; C524898:PLAUR protein, human; D016372:RNA, Antisense; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D010960:Plasminogen Activators; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; D017364:Collagenases; D020781:Matrix Metalloproteinase 1