Induction of the Hyaluronic Acid-Binding Protein, Tumor Necrosis Factor-Stimulated Gene-6, in Cervical Smooth Muscle Cells by Tumor Necrosis Factor-α and Prostaglandin E2

Abstract

Immediately before parturition the cervix undergoes striking changes in structure (ripening) that facilitate dilatation and effacement. Cervical ripening shares many features in common with inflammation-associated tissue remodeling, making it a valuable process to explore with respect to the biochemical events in extracellular matrix restructuring. Cervical ripening can be pharmacologically induced with prostaglandin E(2) (PGE(2)). Among the biochemical changes in the cervix at parturition is a marked increase in the hyaluronic acid (HA) content. HA and HA-binding proteins have been implicated in tissue hydration, release of collagenase, and leukocyte migration, but their roles in cervical ripening have not been explored. In the present study we examined the ability of PGE(2) to induce expression of the HA-binding protein, tumor necrosis factor-stimulated gene (TSG)-6, in human cervical smooth muscle cells (hCSMCs) and compared the PGE(2) response to that of tumor necrosis factor-alpha (TNF-alpha), an established inducer of TSG-6. TNF-alpha stimulated TSG-6 mRNA accumulation in a dose- and time-dependent manner, with the maximal response observed at 10 ng/ml after 6 hours of incubation. PGE(2) stimulated TSG-6 mRNA expression, but the magnitude of response was substantially less than that produced by TNF-alpha, and it was maximal only after 24 hours of incubation. Quantitative real-time polymerase chain reaction was performed to assess the induction of TSG-6 mRNA and nascent transcripts at 24 hours of treatment. Induction of TSG-6 mRNA and nascent transcripts in response to 10 micromol/L of PGE(2) was 5.7-fold and 6.3-fold greater than control values, respectively, whereas TNF-alpha (10 ng/ml) induced TSG-6 mRNA and nascent transcripts by 80-fold and 134-fold, respectively. TNF-alpha and PGE(2) stimulated secretion of TSG-6 into the culture medium as detected by Western blotting. The effects of PGE(2) on secretion of TSG-6 were delayed compared to TNF-alpha. A 1.3-kb fragment of the human TSG-6 proximal promoter drove luciferase expression in transfected hCSMCs. PGE(2) increased TSG-6 promoter activity 1.75-fold. Paradoxically, TNF-alpha reduced TSG-6 promoter activity by 50%. We conclude that hCSMCs express the hyaladherin TSG-6; that TSG-6 expression in these cells is regulated by PGE(2) as well as proinflammatory cytokines; responses of hCSMCs to TNF-alpha and PGE(2) are distinct in terms of magnitude and the time course; and PGE(2) and TNF-alpha exert different effects on the TSG-6 proximal promoter.

TSG-6: a multifunctional protein associated with inflammation

“…1). TSG-6 mRNA and protein are also produced in cumulus oocyte complexes (COCs) following the induction of ovulation (Fülöp et al, 1997;Yoshioka et al, 2000;Carrette et al, 2001;Mukhopadhyay et al, 2001) and by cervical SMCs (cSMCs) treated with PGE2, which promotes cervical ripening (Fujimoto et al, 2002) IαI-TSG-6 Fig. 2.…”

“…This suggests that several distinct pathways regulate TSG-6 expression. For example, cycloheximide does not inhibit TSG-6 transcription in response to TNF, IL-1 or PGE2 (Feng and Liau, 1993;Lee et al, 1993a;Fujimoto et al, 2002) but abrogates growth-factor-induced expression of TSG-6 (Feng and Liau, 1993;Ye et al, 1997). Furthermore, although the upregulation of TSG-6 in response to most stimuli is rapid and relatively short lived, some factors [e.g.…”

“…1) (reviewed in Wisniewski and Vilcek, 1997). It is also becoming clear that TSG-6 is produced in inflammation-like processes, such as ovulation (Fülöp et al, 1997) and cervical ripening (Fujimoto et al, 2002), where its expression is probably induced by prostaglandin E2 (PGE2) (Fujimoto et al, 2002;Ocshner et al, 2003). Growth factors [e.g.…”

The role of cytokines and adhesion molecules in maternal recognition and establishment of pregnancy in pig

“…At this time, the formation of conceptus-uterine extracellular matrix is essential. Tumor necrosis factor stimulated gene (TSG)-6 is induced not only by tumor necrosis factor (TNF-α) but also by IL-1β (Lee et al, 1992) and prostaglandin E2 (PGE) (Fujimoto et al, 2002). Beside the antiinflamatory action (Wisniewski et al, 1996) TSG-6, is thought to be involved in cumulus-oocyte matrix formation in mice because of its ability to bind to and stabilize the covalent bond between the hyaluronic acid and the heavy chain of inter-α-trypsin inhibitor (Richards et al, 2002).…”

TNF?-stimulated gene product (TSG-6) and its binding protein, I?I, in the human intervertebral disc: new molecules for the disc

“…TNFa-stimulated gene-6 (TSG-6) is a 35kD glycoprotein that is considered to be a marker of inflammation [20,33]. It is normally absent or produced at very low levels by cells of mesenchymal origin, but they can be induced to secrete TSG-6 when stimulated by TNFa or other proinflammatory mediators such as prostaglandin E 2 and IL-1, both in vitro and in vivo [5,16,20]. Furthermore, TSG-6 is found in synovial fluid and joint tissues of patients with rheumatoid arthritis (RA) or osteoarthritis (OA) but not in unaffected individuals [2,30].…”