Molecular mechanisms regulating the production of collagenase and TIMP in U937 cells: Evidence for involvement of delayed transcriptional activation and enhanced mRNA stability

Abstract

We have used the human promonocyte-like U937 cell line as a model to study the regulation of interstitial collagenase and tissue inhibitor of metalloproteinases (TIMP) during mononuclear phagocyte development. Our results show that differentiation of U937 cells with exposure to 12-O-tetradecanoylphorbol 13-acetate (TPA) induces a temporally delayed (16-24 h) but marked increase in the biosynthesis and secretion of interstitial collagenase and TIMP. Similarly, steady-state mRNA levels for both proteins rose dramatically during the period of exposure, but again after considerable time delay (12-16 h). For interstitial collagenase, induction was transcriptionally regulated as demonstrated by nuclear run-on experiments, and required the synthesis of proteins as indicated by cycloheximide treatment. However, transcriptional activation of collagenase was never observed prior to 10-12 h; since c-fos is rapidly induced in U937 cells and largely disappears by 2 h (Mitchell et al., 1985), our data strongly suggest that collagenase induction in this system cannot be explained simply or entirely by an AP-1-dependent mechanism. Although TIMP steady-state mRNA levels also increased substantially with cellular differentiation, no transcription was detected by run-on experiments. However, TPA exposure markedly prolonged the half-life of TIMP mRNA from 4 h to > 20 h. While cycloheximide treatment completely blocked TPA-mediated induction of collagenase mRNA, it only marginally interfered with simultaneously induced TIMP mRNA levels. Our results demonstrate that differentiation of U937 monocytic cells is accompanied by markedly enhanced production of both interstitial collagenase and TIMP. However, there are multiple, and perhaps differing, molecular mechanisms regulating these responses.