NF-kappaB-dependent synergistic regulation of CXCL10 gene expression by IL-1beta and IFN-gamma in human intestinal epithelial cell lines.
NF-kappaB-dependent synergistic regulation of CXCL10 gene expression by IL-1beta and IFN-gamma in human intestinal epithelial cell lines.
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人类肠上皮细胞系中IL-1BETA和IFN-GAMMA对CXCL10基因表达的NF-KAPPAB依赖性协同调节。
DOI：
10.1007/s00384-007-0396-6
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发表时间：
2008-03
影响因子：
2.8
通讯作者：
Raddatz D
中科院分区：
文献类型：
--
作者：
Yeruva S;Ramadori G;Raddatz D
CXCL10 （IP-10）是一种参与募集T细胞和单核细胞的趋化因子，目前对CXCL10的表达和分泌知之甚少。我们旨在研究促炎细胞因子白细胞介素(IL)-1β、干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α在肠上皮细胞系中的表达及其调控。采用实时聚合酶链式反应（PCR）、Northern blotting和酶联免疫吸附试验（ELISA）评估Caco-2、HT-29和DLD1人结肠上皮细胞中CXCL10的表达和分泌动力学，分别用IL-1β、TNF-α、IFN-γ单独或联合处理。瞬时转染TGL-IP10 （CXCL10启动子）和TGL-IP10-κB2突变启动子，并对核因子(NF)-κB进行gelshift和supershift。Real-time pcr和ELISA实验表明，IL-1β是Caco-2细胞系CXCL10信使rna （mRNA）表达和蛋白分泌的最强和最早的诱导剂，而INF-γ具有延迟的动力学。TNF-α或IL-1β与IFN-γ对三种细胞系CXCL10 mRNA表达和蛋白分泌均有较强的协同作用。利用特异性NF-κB抑制剂和NF-κB结合缺陷的CXCL10启动子构建体进行的实时PCR和ELISA实验表明，IL-1β对CXCL10的诱导及其与IFN-γ的协同作用依赖于NF-κB。这些数据表明，在结肠上皮细胞中，根据细胞背景和利用NF-κB途径，IL-1β单独和/或与IFN-γ协同作用可能在诱导CXCL10中发挥主要作用。
Little is known about the intestinal epithelial expression and secretion of CXCL10 (IP-10), a chemokine involved in recruiting T cells and monocytes. We aimed to study CXCL10 gene expression and regulation by the pro-inflammatory cytokines interleukin (IL)-1β, interferon (IFN)-γ and tumour necrosis factor (TNF)-α in intestinal epithelial cell lines. CXCL10 expression and secretion kinetics were assessed in Caco-2, HT-29 and DLD1 human colon epithelial cells, treated with IL-1β, TNF-α, IFN-γ alone or in combination with each other by real-time polymerase chain reaction (PCR), Northern blotting and enzyme-linked immunoabsorbent assay (ELISA). Transient transfections with TGL-IP10 (CXCL10 promoter) and TGL-IP10-κB2 mutant promoter and gelshifts and supershifts for nuclear factor (NF)-κB were also performed. Real-time PCRs and ELISA experiments revealed that IL-1β was the strongest and earliest inducer of CXCL10 messenger ribonucleic acid (mRNA) expression and protein secretion in Caco-2 cell line, whereas INF-γ had a delayed kinetics. There was a strong synergistic effect of either TNF-α or IL-1β with IFN-γ both on CXCL10 mRNA expression and protein secretion in all three cell lines. Real-time PCR and ELISA experiments using a specific NF-κB inhibitor and transfection experiments with a NF-κB-binding defective CXCL10 promoter construct revealed that the induction of CXCL10 by IL-1β and its synergism with IFN-γ is NF-κB dependent. These data demonstrate that in colonic epithelial cells, depending on the cellular context and utilizing the NF-κB pathway, IL-1β alone and/or in synergism with IFN-γ may play a major role in the induction of CXCL10.
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