Rapid and transient induction of cyclo-oxygenase 2 by epidermal growth factor in human amnion-derived WISH cells.
Rapid and transient induction of cyclo-oxygenase 2 by epidermal growth factor in human amnion-derived WISH cells.
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在人羊膜来源的 WISH 细胞中，表皮生长因子快速瞬时诱导环加氧酶 2。
DOI：
10.1042/bj3210677
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Kniss,DA
中科院分区：
文献类型：
--
作者：
Perkins,DJ;Kniss,DA
前列腺素(PG)生物合成级联反应的中心酶是PGH2合成酶或环氧合酶(COX)。目前，已鉴定出两种不同的PGH2合成酶/COX亚型：COX-1和COX-2。在许多系统中，COX-1是一种结构性表达的异构体，负责PGs的正常生理产生，而COX-2是一种可诱导的异构体，通过产生高水平的PGs来响应细胞因子、内毒素和生长因子。因此，在表皮生长因子(EGF)刺激的羊膜来源的Wish细胞中，研究了COX-2mRNA和蛋白的调节以及随后PGE2的产生。用浓度为0.01Ő、100 ng/ml的表皮生长因子处理Wish细胞后，COX-2mRNA和蛋白的合成呈剂量依赖性。此外，用10 ng/ml的表皮生长因子刺激Wish细胞后，COX-2mRNA和蛋白的表达在30min内达到稳定水平，然后在2Ő4h内迅速下降至接近基线水平。PGE2的产生也是快速和短暂的。用新型COX-2酶抑制剂NS-398(10~(-5)Ő~(-10)M)预先孵育细胞，可完全抑制PGE_2的形成，并呈剂量依赖关系。然而，地塞米松(地塞米松；0.1ƁM)预先孵育细胞后，在肿瘤坏死因子α(Tumor Numor Fineα，肿瘤坏死因子)刺激的细胞中，PGE2的生成仅减少31%，而PGE2的生物合成却完全减弱。此外，地塞米松(0.1ƁM)仅部分抑制表皮生长因子诱导的COX-2mRNA和蛋白表达，而地塞米松则完全抑制肿瘤坏死因子-α促进的COX-2mRNA和蛋白表达。由此可见，在Wish细胞中，EGF可诱导COX-2mRNA和蛋白的快速而短暂的表达，并随后产生PGE2。
The central enzyme in the prostaglandin (PG) biosynthetic cascade is PGH2synthase or cyclo-oxygenase (COX). At present, two distinct isoforms of PGH2synthase/COX have been identified: COX-1 and COX-2. In many systems, COX-1 is a constitutively expressed isoform that is responsible for normal physiological production of PGs, whereas COX-2 is an inducible isoform that responds to cytokines, endotoxin and growth factors by producing high levels of PGs. The regulation of COX-2 mRNA and protein, and the subsequent production of PGE2, were therefore examined in amnion-derived WISH cells stimulated with epidermal growth factor (EGF). Treatment of WISH cells with EGF (0.01Ő100 ng/ml) elicited dose-dependent synthesis of COX-2 mRNA and proteinde novo. In addition, stimulation of WISH cells with EGF (10 ng/ml) induced steady-state levels of COX-2 mRNA and protein that appeared within 30 min and then declined rapidly to near baseline levels within 2Ő4 h. In contrast, COX-1 protein was unchanged in response to treatment with EGF. PGE2production was also rapid and transient. Preincubation of cells with the novel COX-2 enzymic inhibitor NS-398 (10-5Ő10-10M) completely prevented PGE2formation in a dose-dependent manner. Preincubation of cells in dexamethasone (Dex; 0.1ƁM), however, resulted in only a 31% decrease in PGE2formation in response to EGF (10 ng/ml) while completely attenuating PGE2biosynthesis in tumour necrosis factor α (TNF-α)-stimulated cells. In addition, Dex (0.1ƁM) was only partly effective at preventing EGF-induced COX-2 mRNA and protein expressionde novo, whereas Dex completely inhibited TNF-α-promoted COX-2 mRNA and protein expression. Thus the results presented here demonstrate that EGF induces the rapid but transient expression of COX-2 mRNA and protein and the subsequent production of PGE2in WISH cells.