Abstract
The effect of basement membrane components (laminin, fibronectin and type IV collagen) and lung fibroblasts on type IV collagenase and plasminogen activator activity was investigated in a primary HSV-2-induced hamster fibrosarcoma, and its in vivo derived sublines and in vitro derived clones of varying metastatic potential. Fibronectin and type IV collagen were ineffective at influencing the expression of either type IV collagenase or plasminogen activator activity. Laminin, however, at concentrations of 1-10 micrograms ml-1 added to the serum-free culture supernatants, increased the release of type IV collagenase by up to 100% for the parental cell line. Three highly metastatic sublines (two from in vivo origin and one from in vitro cloning) showed increases of up to 300%. Non-metastatic sublines (two from in vivo origin and one from in vitro cloning), however, showed no increase in type IV collagenase activity. Plasminogen activator release from either the parental line cell or its metastatic sublines and clones, was unaffected by the addition of laminin. Addition of tumour cells to lung fibroblast monolayers resulted in an increased expression of PA activity in the supernatant, whilst type IV collagenase activity was reduced.
Highlights
Type IV collagenase activity was increased up to 100% for the parent cell line in the presence of laminin
Precoating tissue culture flasks with fibronectin or type IV collagen was ineffective at altering the expression of type IV collagenase
Increases in type IV collagenase activity of up to 300% were obtained in the presence of laminin but fibronectin and type IV collagen had no effect (Figure 1)
Summary
AnimalsMale, Syrian golden hamsters, aged between 6 and 10 weeks and weighing 60-90 grams were used in all experiments. The animals were obtained from a closed randomly bred colony at the University of Sheffield and have previously been shown to be syngeneic by skin grafting experiments (Potter & Jennings, unpublished) and mixed lymphocyte reactions in vitro (Teale, unpublished). The four sublines met B, met C, met F and met G were derived from lung nodules in hamsters whose primary parent load had previously been resected; following in vivo passage, in vitro cultures were established and used within 10 passages. Clones S4A and S9E were obtained following double cloning of the parental cell line by the limiting dilution method (Teale & Rees, 1987).
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