雷公藤内酯醇可下调 SK-MEL-2 人黑色素瘤细胞中人 GD3 合酶（hST8Sia I）基因的表达。
Triptolide downregulates human GD3 synthase (hST8Sia I) gene expression in SK-MEL-2 human melanoma cells.
机构信息
Department of Biotechnology, Brain Korea 21 Center for Silver-Bio Industrialization, Dong-A University, Busan 604-714, Korea.
出版信息
Exp Mol Med. 2010 Dec 31;42(12):849-55. doi: 10.3858/emm.2010.42.12.088.
Abstract
In this study, we have shown that gene expression of human GD3 synthase (hST8Sia I) is suppressed by triptolide (TPL) in human melanoma SK-MEL-2 cells. To elucidate the mechanism underlying the downregulation of hST8Sia I gene expression in TPL-treated SK-MEL-2 cells, we characterized the TPL-inducible promoter region within the hST8Sia I gene using luciferase constructs carrying 5'-deletions of the hST8Sia I promoter. Functional analysis of the 5'-flanking region of the hST8Sia I gene demonstrated that the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-κB, functions as the TPL-inducible promoter of hST8Sia I in SK-MEL-2 cells. Site-directed mutagenesis and ChIP analysis indicated that the NF-κB binding site at -731 to -722 is crucial for TPL-induced suppression of hST8Sia I in SK-MEL-2 cells. This suggests that TPL induces down-regulation of hST8Sia I gene expression through NF-κB activation in human melanoma cells.
摘要
在这项研究中，我们已经表明三萜内酯（TPL）可抑制人黑色素瘤 SK-MEL-2 细胞中人类 GD3 合酶（hST8Sia I）的基因表达。为了阐明 TPL 处理的 SK-MEL-2 细胞中 hST8Sia I 基因表达下调的机制，我们使用携带 hST8Sia I 启动子 5'缺失的荧光素酶构建体来表征 hST8Sia I 基因中的 TPL 诱导启动子区域。hST8Sia I 基因 5'侧翼区的功能分析表明，包含转录因子 c-Ets-1、CREB、AP-1 和 NF-κB 结合位点的-1146 至-646 区域在 SK-MEL-2 细胞中作为 hST8Sia I 的 TPL 诱导启动子发挥作用。定点诱变和 ChIP 分析表明，-731 至-722 处的 NF-κB 结合位点对于 TPL 诱导的 SK-MEL-2 细胞中 hST8Sia I 的抑制至关重要。这表明 TPL 通过人黑素瘤细胞中 NF-κB 的激活诱导 hST8Sia I 基因表达的下调。
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