Regulation of the cd38 promoter in human airway smooth muscle cells by TNF-alpha and dexamethasone.
Regulation of the cd38 promoter in human airway smooth muscle cells by TNF-alpha and dexamethasone.
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DOI：
10.1186/1465-9921-9-26
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发表时间：
2008-03-14
影响因子：
5.8
通讯作者：
Kannan MS
中科院分区：
文献类型：
--
作者：
Tirumurugaan KG;Kang BN;Panettieri RA;Foster DN;Walseth TF;Kannan MS
CD 38在人气道平滑肌（HASM）细胞中表达，调节细胞内钙，并且其表达被肿瘤坏死因子α（TNF-α）增强。CD 38在气道高反应性（哮喘的标志）中起作用，因为与野生型小鼠相比，在用细胞因子如IL-13和TNF-α鼻内激发后，缺陷型小鼠的气道高反应性减弱。HASM细胞中CD 38表达的调节涉及转录因子NF-κB B，糖皮质激素通过NF-κ B依赖性和非依赖性机制抑制这种表达。在这项研究中，我们确定是否在HASM细胞中的CD 38表达的转录调控涉及该基因的启动子区域内的响应元件。我们将人cd 38基因的一个假定的3 kb启动子片段克隆到pGL 3基础载体中，位于荧光素酶报告基因的前面。对cd 38启动子区的序列分析表明，该基因含有一个NF-κB和多个AP-1及糖皮质激素反应元件（GRE）基序。用3 kb启动子、缺少NF-κB和一些AP-1位点的1.8 kb截短启动子或具有NF-κB和/或AP-1位点突变的启动子转染HASM细胞。使用电泳迁移率变动分析，我们确定了核蛋白与编码推定的cd 38 NF-κB、AP-1和GRE位点的寡核苷酸的结合，并通过使用适当抗体的凝胶超移分析证实了这种结合的特异性。TNF-α转染HASM细胞后，可诱导3 kb启动子激活2倍。在用1.8 kb启动子或缺乏NF-κB和/或AP-1位点的启动子构建体转染的细胞中，或在存在地塞米松的情况下，在存在TNF-α的情况下没有诱导。TNF-α可增加核蛋白与编码推定的cd 38 NF-κB位点和6个AP-1位点中的一些位点的寡核苷酸的结合，地塞米松可增加核蛋白与某些推定的cd 38 GRES的结合。EMSA结果和cd 38启动子-报告基因分析证实了cd 38启动子中NF-κB、AP-1和GRES在CD 38转录调控中的功能作用。
CD38 is expressed in human airway smooth muscle (HASM) cells, regulates intracellular calcium, and its expression is augmented by tumor necrosis factor alpha (TNF-α). CD38 has a role in airway hyperresponsiveness, a hallmark of asthma, since deficient mice develop attenuated airway hyperresponsiveness compared to wild-type mice following intranasal challenges with cytokines such as IL-13 and TNF-α. Regulation of CD38 expression in HASM cells involves the transcription factor NF-κB, and glucocorticoids inhibit this expression through NF-κB-dependent and -independent mechanisms. In this study, we determined whether the transcriptional regulation of CD38 expression in HASM cells involves response elements within the promoter region of this gene. We cloned a putative 3 kb promoter fragment of the human cd38 gene into pGL3 basic vector in front of a luciferase reporter gene. Sequence analysis of the putative cd38 promoter region revealed one NF-κB and several AP-1 and glucocorticoid response element (GRE) motifs. HASM cells were transfected with the 3 kb promoter, a 1.8 kb truncated promoter that lacks the NF-κB and some of the AP-1 sites, or the promoter with mutations of the NF-κB and/or AP-1 sites. Using the electrophoretic mobility shift assays, we determined the binding of nuclear proteins to oligonucleotides encoding the putative cd38 NF-κB, AP-1, and GRE sites, and the specificity of this binding was confirmed by gel supershift analysis with appropriate antibodies. TNF-α induced a two-fold activation of the 3 kb promoter following its transfection into HASM cells. In cells transfected with the 1.8 kb promoter or promoter constructs lacking NF-κB and/or AP-1 sites or in the presence of dexamethasone, there was no induction in the presence of TNF-α. The binding of nuclear proteins to oligonucleotides encoding the putative cd38 NF-κB site and some of the six AP-1 sites was increased by TNF-α, and to some of the putative cd38 GREs by dexamethasone. The EMSA results and the cd38 promoter-reporter assays confirm the functional role of NF-κB, AP-1 and GREs in the cd38 promoter in the transcriptional regulation of CD38.
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影响因子：
3.6
作者：
Dogan, S;White, TA;Kannan, MS
通讯作者：
Kannan, MS
DOI：
10.1165/ajrcmb.26.4.4681
发表时间：
2002-04-01
影响因子：
6.4
作者：
Ammit, AJ;Lazaar, AL;Panettieri, RA
通讯作者：
Panettieri, RA
影响因子：
7.3
作者：
Barnes, PJ
通讯作者：
Barnes, PJ
影响因子：
4.1
作者：
Dogan, S;Deshpande, DA;Kannan, MS
通讯作者：
Kannan, MS
影响因子：
56.9
作者：
AUPHAN, N;DIDONATO, JA;KARIN, M
通讯作者：
KARIN, M