Transforming Growth Factor-β- and Tumor Necrosis Factor-α-mediated Induction and Proteolytic Activation of MMP-9 in Human Skin

Abstract

Both cytokines and matrix metalloproteinases (MMPs) are active during physiologic and pathologic processes such as cancer metastasis and wound repair. We have systematically studied cytokine-mediated MMP regulation. Cytokine-mediated proteinase induction and activation were initially investigated in organ-cultured human skin followed by determination of underlying cellular and molecular mechanisms using isolated skin cells. In this report we demonstrate that tumor necrosis factor-␣ (TNF-␣) and transforming growth factor-␤ (TGF-␤) synergistically induce pro-MMP-9 in human skin as well as isolated dermal fibroblasts and epidermal keratinocytes. Furthermore, TNF-␣ promotes proteolytic activation of pro-MMP-9 by conversion of the 92-kDa pro-MMP-9 to the 82-kDa active enzyme. This activation occurred only in skin organ culture and not by either isolated fibroblasts or keratinocyte, although the pro-MMP-9 activation could be measured in a cellfree system derived from TNF-␣-activated skin. The cytokine-mediated induction of pro-MMP-9 in dermal fibroblasts was evident by increased mRNA. At the transcription level, we examined the cytokine-mediated transactivation of the 5-region promoter of the human MMP-9 in dermal fibroblasts. The results demonstrated that TNF-␣ and TGF-␤ could independently stimulate the 5-flanking 670-base pair promoter. A TGF-␤-response element (؊474) and an NF-B-binding site (؊601) were identified to be the cis-elements for TGF-␤ or TNF-␣ activation, respectively. Taken together, these findings suggest a specific mechanism whereby multiple cytokines can regulate MMP-9 expression/activation in the cells of human skin. These results imply roles for these cytokines in the regulation of MMP-9 in physiologic and pathologic tissue remodeling.

RGTA OTR4120, a heparan sulfate mimetic, is a possible long‐term active agent to heal burned skin

“…42 Moreover, TGF-b1 contributes to stimulate MMP-9 expression. 43 Thus, OTR4120 may also increase MMP-2 and MMP-9 expression at cutaneous burn sites via a similar TGFb1-related pathway.…”

Interleukin-1α–induced proteolytic activation of metalloproteinase-9 by human skin

“…23 In brief, normal human skin was obtained from the discarded tissue of patients undergoing reconstructive or aesthetic surgery; this procedure was approved by the University of Southern California Institutional Review Board #012022. Full-thickness skin was decontaminated by incubation in 200 U/mL penicillin G sodium, 200 U/mL streptomycin sulfate, and 0.5 mg/mL amphotericin B in Dulbecco modified Eagle medium (DMEM) for 14 hours at 4°C.…”

“…As shown previously, the pro-MMP-9-converting enzyme activity is mostly retained in the Triton-insoluble fraction of skin tissue. 23 To assay the converting activity, we incubated 80 μL of the precleaned conditioned medium or the suspension of tissue fractions with 15 μL of purified pro-MMP-9 together with 5 μL 100 mmol/L CaCl 2 . After 16 hours of incubation at 37°C, the incubated samples were resolved by gelatinolytic zymography to measure relative amounts of active and pro-MMP-9.…”

PMA-induced up-regulation of MMP-9 is regulated by a PKCα-NF-κB cascade in human lung epithelial cells

“…Previous work has suggested that the induction of MMP-9 is affected by various factors such as proinflammatory cytokines, TGF-, EGF, and certain oncogenes (Sehgal and Thompson, 1999;Han et al, 2001;Nee et al, 2004;Wu et al, 2004;Chou et al, 2006). Phorbol myristate acetate (PMA), a phorbol ester, is known to substitute for DAG as a high affinity ligand for conventional protein kinase C (PKC) and novel PKC isoforms (Easom et al, 1989).…”