NF-κB Enhances Androgen Receptor Expression through 5′-UTR Binding in Gingival Cells
NF-κB Enhances Androgen Receptor Expression through 5′-UTR Binding in Gingival Cells
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DOI：
10.1177/0022034515594117
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发表时间：
2015-10-01
影响因子：
7.6
通讯作者：
Lu, H. K.
中科院分区：
文献类型：
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作者：
Chang, J. H.;Wang, L. F.;Lu, H. K.
二氢吡啶诱导的牙龈过度生长(DIGO)是一种在高血压患者治疗中观察到的副作用。这种疾病因炎症而加重。硝苯地平(NIF)是一种二氢吡啶，通过增加雄激素受体(AR)的表达而导致牙龈过度生长。此外，促炎细胞因子白介素1(IL-1)通过AR诱导DIGO成纤维细胞表达I型胶原。这些观察结果促使我们研究核因子kappaB(NF-B)是否以及如何影响DIGO中AR的表达。因此，从DIGO患者和健康人的组织中获得的牙龈成纤维细胞被IL-1、NIF或两者联合刺激。用实时荧光定量聚合酶链式反应和Western blotting检测基因和蛋白的表达。两种药物诱导的AR、结缔组织生长因子和I型胶原的mRNA表达均呈高度相关。Western印迹分析显示，IL-1和NIF在DIGO细胞中比在正常细胞中更能增加和激活核因子-B。凝胶迁移率改变分析表明，AR基因的启动子和5-非翻译区(5-UTRs)含有3个与NF-B p65亚基结合的位点。染色质免疫沉淀分析显示，牙周成纤维细胞中的核因子-B p65亚基与AR 5-UTRs相关。定点突变研究表明，核因子-B结合位点的突变降低了NIF和IL-1诱导的AR启动子的活性。综上所述，这些数据表明，核因子-B是AR基因表达的重要转录调节因子，因此在DIGO成纤维细胞的胶原过度生产中发挥关键作用。
Dihydropyridine-induced gingival overgrowth (DIGO) is a side effect observed in patients treated for hypertension. The disease is aggravated by inflammation. Nifedipine (Nif), a dihydropyridine, causes gingival overgrowth by increasing the expression of the androgen receptor (AR). Furthermore, the proinflammatory cytokine interleukin 1 (IL-1) induces collagen 1(I) expression through the AR in DIGO fibroblasts. These observations prompted us to investigate whether and how nuclear factor kappa B (NF-B) affects AR expression in DIGO. Therefore, gingival fibroblasts obtained from the tissues of patients with DIGO and healthy subjects were stimulated with IL-1, Nif, or both. mRNA and protein expression was detected with real-time polymerase chain reaction and Western blotting. High correlation coefficients were observed for the mRNA expression of the AR, connective tissue growth factor, and collagen 1(I) induced by both drugs. Western blot analysis showed that IL-1 and Nif increased and activated NF-B more in DIGO cells than in healthy cells. An electrophoretic mobility shift assay demonstrated that the promoter and 5-untranslated regions (5-UTRs) of the AR gene contains 3 binding sites for the NF-B p65 subunit. A chromatin immunoprecipitation assay revealed that the NF-B p65 subunit was associated with AR 5-UTRs in gingival fibroblasts. A site-directed mutagenesis study indicated that a mutation of NF-B binding sites reduced Nif- and IL-1-induced AR promoter activities. Collectively, these data indicate that NF-B is an essential transcriptional regulator of AR gene expression and thus plays a crucial role in collagen overproduction in DIGO fibroblasts.