Biotechnology Research Division, National Fisheries Research and Development Institute (NFRDI), 408-1 Sirang-ri, Gijang-up, Gijang-gun, Busan 619-705, Republic of Korea.
Fish Shellfish Immunol. 2010 May-Jun;28(5-6):961-4. doi: 10.1016/j.fsi.2010.01.014. Epub 2010 Feb 6.
Interleukin (IL)-6 plays important roles in the regulation of the immune response and inflammation in many cell types and its induction by bacterial endotoxins or cytokines is regulated at the transcriptional level. The present study demonstrates the isolation and characterization of the flounder IL-6 promoter sequence and its transcriptional regulation in olive flounder (hirame) natural embryo (HINAE) cells. The promoter region (-400 to +79 bp from the transcription initiation site) of the flounder IL-6 gene contains putative cis-regulatory elements for CCAAT-enhancer-binding proteins (C/EBP; -346 to -355 bp and -166 to -160 bp), cAMP response element binding (CREB; -81 to -85 bp), the activator protein 1 (AP-1; -56 to -62 bp), and NF-kappaB (-39 to -48 bp). Transfection of p65 stimulated the PoIL-6-luc-WT reporter, but not the PoIL-6-luc-kappaB mt reporter, and treatment with LPS augmented p65-stimulated reporter activity in HINAE cells. In contrast, transfection of C/EBPbeta or c-Jun failed to induce synergistic effects in the LPS-driven PoIL-6-luc-WT reporter activity. These results give us new insight into the regulation of flounder IL-6 transcription by the p65 NF-kappaB subunit.
白细胞介素 (IL)-6 在许多细胞类型的免疫反应和炎症调节中发挥重要作用，其诱导作用由细菌内毒素或细胞因子在转录水平上调节。本研究展示了牙鲆 IL-6 启动子序列的分离和特征及其在牙鲆自然胚胎 (HINAE) 细胞中的转录调控。牙鲆 IL-6 基因的启动子区域（从转录起始位点起-400 至+79bp）包含 CCAAT 增强子结合蛋白 (C/EBP；-346 至-355bp 和-166 至-160bp)、cAMP 反应元件结合蛋白 (CREB；-81 至-85bp)、激活蛋白 1 (AP-1；-56 至-62bp) 和 NF-κB(-39 至-48bp) 的顺式调控元件。p65 的转染刺激了 PoIL-6-luc-WT 报告基因，但不刺激 PoIL-6-luc-kappaB mt 报告基因，而 LPS 的处理增强了 HINAE 细胞中 p65 刺激的报告基因活性。相比之下，C/EBPβ或 c-Jun 的转染未能在 LPS 驱动的 PoIL-6-luc-WT 报告基因活性中诱导协同作用。这些结果使我们对 p65 NF-κB 亚基调节牙鲆 IL-6 转录有了新的认识。
