Differential regulation of collagenase and stromelysin mRNA in late passage cultures of human fibroblasts.

Nontransformed human fibroblast cell cultures have been extensively studied as an in vitro model for cellular senescence. Recently there has been considerable interest in using the human fibroblast in the identification of genes relevant to the process of replicative senescence. We demonstrated that in comparison with early passage cultures the expression of collagenase and stromelysin mRNAs and proteins was increased > 8 x in late passage cultures of human fibroblasts and, in addition, expression of Il-1 alpha, a cytokine that regulates collagenase and stromelysin expression, was also significantly increased in late passage cell cultures. These findings suggested the hypothesis that constitutive Il-1 alpha expression in late passage cells may coordinately regulate the age-associated increase in the expression of collagenase and stromelysin. To test this hypothesis we examined the effects of long-term Il-1 alpha treatment, serum starvation, and cycloheximide inhibition on collagenase and stromelysin mRNA levels in early and late passage human fibroblast cell cultures. Here we report that in late passage cell cultures, collagenase and stromelysin mRNAs respond differentially to Il-1 alpha, serum starvation, and cycloheximide addition. Continuous exposure to Il-1 alpha reduced the half-life of stromelysin mRNA but had little effect on the half-life of collagenase mRNA. In contrast to stromelysin, the collagenase mRNA level is dependent on serum factors. Collagenase is induced during recovery from cycloheximide inhibition, but stromelysin expression is not affected. These results establish that collagenase and stromelysin mRNAs are differentially regulated in both early and late passage human fibroblasts and suggest that the mechanisms responsible for the age-associated increase in the two mRNAs are different. In addition, these studies support the conclusion that continuous long-term exposure to Il-1 alpha, a condition that is characteristic of late passage cells, is not the factor responsible for the high levels of collagenase expression, but may be critical for stromelysin expression.