Proinflammatory cytokines induce amelotin transcription in human gingival fibroblasts
Proinflammatory cytokines induce amelotin transcription in human gingival fibroblasts
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DOI：
10.2334/josnusd.56.261
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发表时间：
2014-12-01
影响因子：
1.9
通讯作者：
Ogata, Yorimasa
中科院分区：
文献类型：
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作者：
Nakayama, Yohei;Takai, Hideki;Ogata, Yorimasa
AMTN是一种分泌性蛋白，主要在牙釉质形成成熟阶段转录，并定位于接合上皮。我们研究了慢性牙周炎患者非炎症牙龈和炎症牙龈之间AMTN基因表达水平的差异。从这些组织中分离总rna，并通过DNA微阵列监测其基因表达谱。实时荧光定量PCR证实AMTN mRNA在炎症牙龈和培养的人牙龈成纤维细胞（HGF）中的诱导作用。使用与荧光素酶报告基因连接的小鼠AMTN基因启动子片段的嵌合结构进行瞬时转染试验。用免疫组化方法评估炎症和非炎症牙龈中AMTN的定位。在众多差异表达基因中，AMTN mRNA水平在炎症牙龈中显著升高。用白细胞介素-1 β (IL-1 β)、IL-6和肿瘤坏死因子- α (tnf - α)处理HGF可诱导AMTN mRNA的表达，并增加AMTN启动子构建体的荧光素酶活性。炎症牙龈结缔组织和结膜上皮中检测到AMTN蛋白。这些发现表明，促炎细胞因子诱导AMTN基因在人牙龈成纤维细胞中的表达，并提示AMTN在牙龈炎症中的作用。
Amelotin (AMTN) is a secreted protein transcribed predominantly during the maturation stage of enamel formation and localized in the junctional epithelium. We investigated differences in the levels of AMTN gene expression between non-inflamed gingiva and inflamed gingiva from patients with chronic periodontitis. Total RNAs were isolated from these tissues and their gene expression profiles were monitored by DNA microarray. The observed induction of AMTN mRNA in inflamed gingiva and cultured human gingival fibroblasts (HGF) was confirmed by real-time PCR. Transient transfection assays were performed using chimeric constructs of mouse AMTN gene promoter fragments linked to a luciferase reporter gene. Immunohistochemical localization of AMTN in inflamed and non-inflamed gingiva was assessed by immunohistochemistry. Among many differentially expressed genes, the level of AMTN mRNA was significantly increased in inflamed gingiva. Treatment of HGF with interleukin-1 beta (IL-1 beta), IL-6 and tumor necrosis factor-alpha (TNF-alpha) induced the expression of AMTN mRNA, and increased the luciferase activities of the AMTN promoter constructs. AMTN protein was detected in inflamed gingival connective tissue and junctional epithelium. These findings demonstrate that proinflammatory cytokines induce AMTN gene expression in human gingival fibroblasts and suggest a role for AMTN in gingival inflammation.