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
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
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DOI：
10.1016/s0002-9440(10)62575-8
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发表时间：
2002-04-01
影响因子：
6
通讯作者：
Strauss, JF
中科院分区：
文献类型：
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作者：
Fujimoto, T;Savani, RC;Strauss, JF
就在分娩前，宫颈结构发生了显著的变化(成熟)，这有利于扩张和消退。宫颈成熟与炎症相关的组织重塑有许多共同的特征，这使得它成为一个有价值的过程，关于细胞外基质重塑中的生化事件。前列腺素E-2(PGE(2))可在药理上诱导宫颈成熟。在分娩时宫颈的生化变化中，透明质酸(HA)含量显著增加。透明质酸和透明质酸结合蛋白与组织水合、胶原酶的释放和白细胞的迁移有关，但它们在宫颈成熟中的作用尚不清楚。在本研究中，我们检测了PGE(2)诱导人宫颈平滑肌细胞(HCSMCs)HA结合蛋白-肿瘤坏死因子刺激基因(TSG)-6表达的能力，并比较了PGE(2)与TSG-6的诱导剂肿瘤坏死因子-α(TNF-α)的反应。肿瘤坏死因子-α以剂量和时间依赖的方式刺激TSG-6mRNA的表达，孵育6h后最大反应浓度为10 ng/ml。PGE(2)可刺激TSG-6mRNA的表达，但其反应幅度明显小于肿瘤坏死因子-α，且仅在孵育24小时后达到最大。用实时定量聚合酶链式反应检测治疗24小时后TSG-6mRNA和新生转录产物的诱导。PGE(2)对TSG-6mRNA和新生转录产物的诱导作用分别是对照组的5.7倍和6.3倍，而肿瘤坏死因子-α(10 ng/ml)诱导的TSG-6mRNA和新生转录产物分别是对照组的80倍和134倍。Western blotting检测，肿瘤坏死因子-α和前列腺素E(2)刺激培养上清液中TSG-6的分泌。与肿瘤坏死因子-α相比，前列腺素E(2)对TSG-6分泌的影响延迟。人TSG-6近端启动子的1.3 kb片段驱动了荧光素酶在hCSMCs中的表达。PGE2使TSG-6启动子活性提高1.75倍。矛盾的是，肿瘤坏死因子-α使TSG-6启动子的活性降低了50%。结论：hCSMCs表达透明粘附素TSG-6；TSG-6表达受PGE(2)和促炎细胞因子的调节；hCSMCs对TNF-α和PGE(2)的反应在幅度和时程上不同；PGE(2)和TNF-α对TSG-6近端启动子的作用不同。
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 mumol/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. PGE2 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.