Interleukin-1 beta and transforming growth factor-alpha/epidermal growth factor induce expression of M(r) 95,000 type IV collagenase/gelatinase and interstitial fibroblast-type collagenase by rat mucosal keratinocytes

• Published in: The Journal of biological chemistry Vol. 268; no. 25; pp. 19143 - 19151
• Main Authors: Lyons, J.G., Birkedal-Hansen, B., Pierson, M.C., Whitelock, J.M., Birkedal-Hansen, H.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.09.1993; American Society for Biochemistry and Molecular Biology
• Subjects: Alkaloids - pharmacology; Animals; Cell Line; Collagenases - metabolism; Dexamethasone - pharmacology; Endopeptidases - metabolism; Enzyme Induction; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor - pharmacology; Fibroblasts - enzymology; Gelatinases; Interleukin-1 - pharmacology; Keratinocytes - enzymology; Mammary Neoplasms, Experimental; Molecular Weight; Mouth Mucosa - enzymology; Protein Kinase C - metabolism; Rats; Staurosporine; Tetradecanoylphorbol Acetate - pharmacology; Transforming Growth Factor alpha - pharmacology; Tumor Cells, Cultured
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)46745-7

Rat mucosal keratinocytes serially propagated under permanently serum-free conditions responded to interleukin (IL)-1 beta/IL-alpha and to transforming growth factor (TGF)-alpha/epidermal growth factor (EGF) (as well as to 12-O-tetradecanoylphorbol-13-acetate (TPA)) by upregulation of M(r) 95,000 gelatinase (MMP-9) (M(r) 95K GL) and fibroblast-type collagenase (MMP-1) (FIB-CL), whereas control cells expressed barely detectable levels of either of these enzymes. The cells secreted 8-10 micrograms/10(6) cells/day (M(r) 95K GL) and 2-3 micrograms/10(6) cells/day (FIB-CL) of enzyme protein for at least 24 h when maximally induced. This level was attained only after a 24-h lag period, and the earliest emergence of enzyme protein in the culture medium required 10-14 h. IL-1 beta was by far the most potent cytokine with maximal effect already at 10(-10) M, whereas IL-1 alpha, TGF-alpha, and EGF required 20-100-fold higher concentrations. Pretreatment of the cells with TPA (10(-7) M) abolished the subsequent response to IL-1 beta, TGF-alpha, and EGF and at the same time resulted in > 90% reduction of cytosolic protein kinase C activity. Surprisingly, staurosporine, a potent kinase inhibitor, not only failed to block growth factor/cytokine responses but itself stimulated expression of the enzymes at a magnitude comparable to TPA. The inducing effect of TGF-alpha/EGF was down-regulated by 70-85% by 10(-7) M dexamethasone. Dexamethasone was less effective in ablating the IL-1 beta response yielding 60% reduction M(r) 95K GL and little or no reduction of FIB-CL. Dexamethasone also failed to block the TPA response.