白细胞介素-6调节人牙龈上皮细胞中ODAM基因的表达。

IF 2.2 4区 医学 Q2 DENTISTRY, ORAL SURGERY & MEDICINE
Zhenyu Jin, Arisa Yamaguchi, Hideki Takai, Yohei Nakayama, Yorimasa Ogata
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引用次数: 0

摘要

目的:牙源性成釉细胞相关蛋白(ODAM)是一种由结上皮(JE)和成熟成釉细胞产生的小分泌蛋白。它在牙形成中起作用,并介导乙脑与牙釉质的粘附。我们利用人牙龈上皮细胞研究白细胞介素(IL)-6对乙脑中ODAM基因表达的调控机制。方法:用IL-6 (10 ng/mL)刺激Ca9-22、Sa3和HSY细胞,提取总RNA和蛋白质。采用实时聚合酶链反应和Western blot方法检测ODAM mRNA和蛋白的表达水平。荧光素酶(LUC)测定采用不同长度的ODAM基因启动子序列的LUC结构。通过凝胶迁移位移和染色质免疫沉淀(ChIP)分析来研究转录因子与基因启动子内应答元件的结合。结果:IL-6处理可提高ODAM mRNA和蛋白的表达。此外,它还诱导了ODAM基因的启动子活性,而LUC活性被蛋白激酶A、酪氨酸激酶、MEK1/2、磷脂酰肌醇3-激酶、核因子-κB、转录信号传导和激活因子(STAT) 3和糖蛋白130的抑制剂抑制。凝胶迁移位移和ChIP分析显示,IL-6诱导阴阳1 (YY1)、CCAAT/增强子结合蛋白(C/EBP) β、GATA结合蛋白(GATA)和phospho-STAT3与YY1、C/EBP、GATA和干扰素γ激活转录元件(GATE) 1-3个元件结合。结论:这些发现表明IL-6通过靶向人ODAM基因启动子区的YY1、C/EBP、GATA和GATE1-3元件上调ODAM基因的表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Interleukin-6 regulates human ODAM gene expression in gingival epithelial cells.

Purpose: Odontogenic ameloblast-associated protein (ODAM) is a small secretory protein produced by the junctional epithelium (JE) and mature ameloblasts. It plays a role in odontogenesis and mediates the adhesion of JE to enamel. We used human gingival epithelial cells to evaluate the mechanism of ODAM gene expression regulation in the JE by interleukin (IL)-6.

Methods: Ca9-22, Sa3, and HSY cells were stimulated with IL-6 (10 ng/mL), after which total RNA and proteins were extracted. Real-time polymerase chain reaction and Western blot analyses were performed to assess the expression levels of ODAM mRNA and protein. Luciferase (LUC) assays were employed using LUC constructs with varying lengths of the ODAM gene promoter sequence. Gel mobility shift and chromatin immunoprecipitation (ChIP) analyses were conducted to investigate the binding of transcription factors to response elements within the gene promoter.

Results: Treatment with IL-6 increased the expressions of ODAM mRNA and protein. Additionally, it induced promoter activity of the ODAM gene, while LUC activity was suppressed by inhibitors of protein kinase A, tyrosine kinase, MEK1/2, phosphatidylinositol 3-kinase, nuclear factor-κB, signal transducer and activator of transcription (STAT) 3, and glycoprotein 130. Gel mobility shift and ChIP analyses revealed that IL-6 induced the binding of yin yang 1 (YY1), CCAAT/enhancer-binding protein (C/EBP) β, GATA binding protein (GATA), and phospho-STAT3 to the YY1, C/EBP, GATA, and interferon-γ activated transcriptional element (GATE) 1-3 elements.

Conclusions: These findings indicate that IL-6 upregulates ODAM gene expression by targeting the YY1, C/EBP, GATA, and GATE1-3 elements in the promoter region of the human ODAM gene.

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来源期刊
Journal of Periodontal and Implant Science
Journal of Periodontal and Implant Science DENTISTRY, ORAL SURGERY & MEDICINE-
CiteScore
3.30
自引率
5.30%
发文量
38
期刊介绍: Journal of Periodontal & Implant Science (JPIS) is a peer-reviewed and open-access journal providing up-to-date information relevant to professionalism of periodontology and dental implantology. JPIS is dedicated to global and extensive publication which includes evidence-based original articles, and fundamental reviews in order to cover a variety of interests in the field of periodontal as well as implant science.
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