Iwai Yasunobu, Noda Keisuke, Yamazaki Mizuho, Mezawa Masaru, Takai Hideki, Nakayama Yohei, Kitagawa Masae, Takata Takashi, Ogata Yorimasa
Department of Periodontology, Nihon University School of Dentistry at Matsudo.
Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo.
J Oral Sci. 2018;60(4):601-610. doi: 10.2334/josnusd.17-0473.
Follicular dendritic cell-secreted protein (FDC-SP) is expressed in FDCs, human periodontal ligament (HPL) cells, and junctional epithelium. To evaluate the effects of interleukin-1 beta (IL-1β) on FDC-SP gene expression in immortalized HPL cells, FDC-SP mRNA and protein levels in HPL cells following stimulation by IL-1β were measured by real-time polymerase chain reaction and Western blotting. Luciferase (LUC), gel mobility shift, and chromatin immunoprecipitation (ChIP) analyses were performed to study the interaction between transcription factors and promoter regions in the human FDC-SP gene. IL-1β (1 ng/mL) induced the expression of FDC-SP mRNA and protein levels at 3 h, and reached maximum levels at 12 h. IL-1β increased LUC activities of constructs (-116FDCSP - -948FDCSP) including the FDC-SP gene promoter. Transcriptional inductions by IL-1β were partially inhibited by 3-base-pair (3-bp) mutations in the Yin Yang 1 (YY1), GATA, CCAAT-enhancer-binding protein2 (C/EBP2), or C/EBP3 in the -345FDCSP. IL-1β-induced -345FDCSP activities were inhibited by protein kinase A, tyrosine-kinase, mitogen-activated protein kinase (MEK)1/2, and PI3-kinase inhibitors. The results of gel shift and ChIP assays revealed that YY1, GATA, and C/EBP-β interacted with the YY1, GATA, C/EBP2, and C/EBP3 elements that were increased by IL-1β. These studies demonstrate that IL-1β increases FDC-SP gene transcription in HPL cells by targeting YY1, GATA, C/EBP2, and C/EBP3 in the human FDC-SP gene promoter.
滤泡树突状细胞分泌蛋白（FDC-SP）在滤泡树突状细胞、人牙周膜（HPL）细胞和结合上皮中表达。为了评估白细胞介素-1β（IL-1β）对永生化HPL细胞中FDC-SP基因表达的影响，通过实时聚合酶链反应和蛋白质免疫印迹法检测了IL-1β刺激后HPL细胞中FDC-SP mRNA和蛋白水平。进行荧光素酶（LUC）、凝胶迁移率变动分析和染色质免疫沉淀（ChIP）分析以研究转录因子与人FDC-SP基因启动子区域之间的相互作用。IL-1β（1 ng/mL）在3小时时诱导FDC-SP mRNA和蛋白水平表达，并在12小时时达到最高水平。IL-1β增加了包含FDC-SP基因启动子的构建体（-116FDCSP - -948FDCSP）的LUC活性。IL-1β的转录诱导在-345FDCSP中的阴阳1（YY1）、GATA、CCAAT增强子结合蛋白2（C/EBP2）或C/EBP3中被3个碱基对（3-bp）突变部分抑制。IL-1β诱导的-345FDCSP活性被蛋白激酶A、酪氨酸激酶、丝裂原活化蛋白激酶（MEK）1/2和PI3激酶抑制剂抑制。凝胶迁移率变动分析和ChIP分析结果显示，YY1、GATA和C/EBP-β与IL-1β增加的YY1、GATA、C/EBP2和C/EBP3元件相互作用。这些研究表明，IL-1β通过靶向人FDC-SP基因启动子中的YY1、GATA、C/EBP2和C/EBP3来增加HPL细胞中FDC-SP基因的转录。
