Transcriptional regulation of β-defensin-2 by lipopolysaccharide in cultured human cervical carcinoma (HeLa) cells
Transcriptional regulation of β-defensin-2 by lipopolysaccharide in cultured human cervical carcinoma (HeLa) cells
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DOI：
10.1016/j.femsim.2005.01.008
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发表时间：
2005-07-01
影响因子：
--
通讯作者：
Takashiba, S
中科院分区：
文献类型：
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作者：
Mineshiba, J;Myokai, F;Takashiba, S
人β-防御素-2(HBD-2)是一种对细菌、酵母菌和真菌具有广谱抗菌活性的抗菌肽。在这里，我们分析了hBD-2在培养的人宫颈癌(HeLa)细胞中的转录调控。HBD-2基因启动子区-329~-39位DNA含有转录因子的共同结合位点，1个为IL-6表达的核因子结合位点，2个为核因子-kappa B的结合位点。启动子活性的报告基因分析表明，该区域对内毒素的反应性最高。此外，两个核因子-kappa B结合位点的突变导致对内毒素的反应性显著降低，而核因子-白介素6结合位点的突变导致基本启动子活性的升高。凝胶迁移率改变分析表明，内毒素诱导HeLa核因子与含有两个核因子-kappaB结合位点的60bp探针结合，提示这两个结合位点是结合所必需的。我们的结果表明，在HeLa细胞中，这两个核因子-kappa B结合位点参与了内毒素介导的HBD-2转录，而核因子-IL6结合位点抑制了非内毒素依赖的HBD-2转录。(C)2005年欧洲微生物学会联合会。爱思唯尔出版，版权所有。
Human beta-defensin-2 (hBD-2) is an antimicrobial peptide with a broad spectrum of antimicrobial activity against bacteria, yeast and fungi. Here, we analyzed the transcriptional regulation of hBD-2 in cultured human cervical carcinoma (HeLa) cells with or without lipopolysaccharide (LPS). DNA from position -329 to -39 in the hBD-2 promoter region contained the consensus binding sites for transcription factors, one site for nuclear factor for IL-6 expression (NF-IL6) and two sites for nuclear factor-kappa B (NF-kappa B). Reporter gene assays for promoter activity revealed that the region had the highest level of responsiveness to LPS. Furthermore, mutations in both of the NF-kappa B binding sites caused a significant reduction of the responsiveness to LPS, whereas mutation in the NF-IL6 binding site resulted in an elevation of the basal promoter activity. Electrophoretic mobility shift assays demonstrated that LPS induced the binding of HeLa nuclear factors to 60-bp probe containing the two NF-kappa B binding sites, suggesting that the sites were essential for the binding. Our results suggest that the two NF-kappa B binding sites contribute to LPS-mediated hBD-2 transcription while the NF-IL6 binding site represses LPS-independent hBD-2 transcription in the HeLa cells. (c) 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.