Synergistic activation of interleukin-8 gene transcription by all-trans-retinoic acid and tumor necrosis factor-alpha involves the transcription factor NF-kappa B
Synergistic activation of interleukin-8 gene transcription by all-trans-retinoic acid and tumor necrosis factor-alpha involves the transcription factor NF-kappa B
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DOI：
10.1074/jbc.271.43.26954
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发表时间：
1996-10-25
影响因子：
4.8
通讯作者：
Lindley, IJD
中科院分区：
文献类型：
--
作者：
Harant, H;deMartin, R;Lindley, IJD
IL-1或肿瘤坏死因子（TNF）对白细胞介素-8（IL-8）的诱导以及干扰素或糖皮质激素对IL-8的抑制已显示涉及5 '侧翼区核苷酸-94和-71之间的序列以及转录因子NF-IL-6和NF-κ B。A3细胞系来源于人黑色素瘤细胞系G-361，通过用与荧光素酶编码区融合的IL-8启动子的一部分（从转录起始的核苷酸-101至+40）稳定转染。这些调控序列足以通过全反式视黄酸（ATRA）、9-顺式视黄酸、IL-1 β或TNF-α进行转录激活。同时用ATRA和TNF-α处理A3细胞导致荧光素酶表达和IL-8 mRNA水平的剂量和时间依赖性协同增加。亲本细胞系的瞬时转染证明NF-κ B结合位点对于这种协同反式激活是必需的。A3细胞核提取物的电泳迁移率变化分析表明，与单独的TNF-α诱导相比，ATRA和TNF-α刺激超过16小时导致NF-κ B结合增强。同时用ATRA和TNF-α处理也导致与IL-8 NF-κ B位点结合的NF-κ B B复合物的组成发生变化，阻止了两种TNF-α诱导的结合活性的形成。我们认为，这些复合物包括抑制因子，当删除，允许增强结合NF-κ B的同源位点。
Induction of interleukin-8 (IL-8) by IL-1 or tumor necrosis factor (TNF), and repression by interferons or glucocorticoids have been shown to involve sequences between nucleotides -94 and -71 of the 5'-flanking region, and the transcription factors NF-IL-6 and NF-kappa B. The A3 cell line was derived from the human melanoma cell line G-361 by stable transfection with part of the IL-8 promoter (nucleotides -101 to +40 from transcription start) fused to the luciferase coding region. These regulatory sequences were sufficient for transcriptional activation by all-trans-retinoic acid (ATRA), 9-cis-retinoic acid, IL-1 beta, or TNF-alpha. Simultaneous treatment of A3 cells with ATRA and TNF-alpha resulted in a dose- and time-dependent synergistic increase in luciferase expression and IL-8 mRNA levels. Transient transfections of the parental cell line demonstrated that the NF-kappa B binding site is essential for this synergistic transactivation. Electrophoretic mobility shift assays with nuclear extracts of A3 cells showed that stimulation with ATRA and TNF-alpha for more than 16 h resulted in enhanced NF-kappa B binding compared to that induced by TNF-alpha alone. The simultaneous treatment with ATRA and TNF-alpha also resulted in changes in the composition of NF-kappa B complexes bound to the IL-8 NF-kappa B site, preventing the formation of two TNF-alpha-inducible binding activities. We suggest that these complexes consist of repressive factors which, when removed, allow enhanced binding of NF-kappa B to its cognate site.