Stimulation of collagenase production by rat osteosarcoma cells can occur in a subpopulation of cells.
Recent studies have indicated that neutral collagenase can be produced in bones of rats. In addition, it has been demonstrated by in vitro studies that the enzyme is likely secreted by osteoblasts. Cells of the osteoblastic tumor cell line UMR-106 can be stimulated to produce not only collagenase, but also collagenase inhibitor and plasminogen activator. However, it is conceivable that not all osteoblasts produce all of these proteins. In this study, in which UMR cells were maximally stimulated with PTH, only a subpopulation of cells was observed to produce enhanced levels of collagenase but all cells had the ability to synthesize plasminogen activator. Cells of the rat osteosarcoma line UMR-106-01 were stained for the presence of collagenase and tissue plasminogen activator using an immunohistochemical procedure. In many cases, the cells were exposed to monensin for the final 3 h of incubation as well as to the inducing agent PTH. Monensin prevented export of the enzymes, enabling them to be visualized within their cell or origin. Maximal stimulation of collagenase was demonstrated to occur 8 h after exposure to 10(-8) -10(-7) M PTH. Under these conditions, 14-17% of the cells appeared to synthesize elevated amounts of collagenase (as determined by intense staining). Without PTH stimulation, there was a low level of collagenase in all cells, but less than 1% of the cells stained heavily for the enzyme. In contrast, strong staining for plasminogen activator was observed in all cells with or without PTH treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
Strege
D W
DW
Department of Orthopedic Surgery, St. Louis University Medical Center, MO 63110.
Kahn
A J
AJ
Jeffrey
J J
JJ
Partridge
N C
NC
D000818:Animals; D002452:Cell Count; D007124:Immunoenzyme Techniques; D003012:Microbial Collagenase / Q000096:biosynthesis*; D010006:Osteoblasts / Q000201:enzymology*; D012516:Osteosarcoma / Q000201:enzymology* / Q000473:pathology; D010281:Parathyroid Hormone / Q000494:pharmacology; D010960:Plasminogen Activators / Q000096:biosynthesis; D051381:Rats; D014407:Tumor Cells, Cultured
D010281:Parathyroid Hormone; D010960:Plasminogen Activators; D003012:Microbial Collagenase