Wang Jianguo, Mahmud Shawn A, Bitterman Peter B, Huo Yuqing, Slungaard Arne
Sections of Hematology, Oncology, and Transplantation, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Vascular Biology Center, University of Minnesota, Minneapolis, Minnesota 55455.
Sections of Medicine Department, University of Minnesota, Minneapolis, Minnesota 55455; Sections of Pulmonary and Critical Care, University of Minnesota, Minneapolis, Minnesota 55455.
J Biol Chem. 2007 Sep 28;282(39):28408-28418. doi: 10.1074/jbc.M703586200. Epub 2007 Aug 3.
Histone deacetylase inhibitors (HDACi), such as trichostatin A (TSA), can regulate gene expression by promoting acetylation of histones and transcription factors. Human tissue factor (TF) expression is partly governed by a unique, NF-kappaB-related "TF-kappaB" promoter binding site. We find that TSA and four other HDACi (apicidin, MS-275, sodium butyrate, and valproic acid) all inhibit by approximately 90% TF activity and protein level induction in human umbilical vein endothelial cells stimulated by the physiologic agonists tumor necrosis factor (TNF)-alpha, interleukin-1beta, lipopolysaccharide, and HOSCN without affecting expression of the NF-kappaB-regulated adhesion molecules ICAM-1 and E-selectin. TSA and butyrate also blunt TF induction approximately 50% in vitro in peripheral blood mononuclear cells and in vivo in thioglycolate-elicited murine peritoneal macrophages. In human umbilical vein endothelial cells, TSA attenuates by approximately 70% TNF-alpha stimulation of TF mRNA transcription without affecting that of ICAM-1. By electrophoretic mobility shift assay analyses, TNF-alpha and lipopolysaccharide induce strong p65/p50 and p65/c-Rel heterodimer binding to both NF-kappaB and TF-kappaB probes. TSA nearly abolishes TF-kappaB binding without affecting NF-kappaB binding. A chromatin immunoprecipitation assay and a promoter-luciferase reporter system confirm that TSA inhibits TF-kappaB but not NF-kappaB activation. Chromatin immunoprecipitation and small interfering RNA inhibitor studies demonstrate that HDAC3 plays a significant role in TNF-alpha-mediated TF induction. Thus, HDACi transcriptionally inhibit agonist-induced TF expression in endothelial cells and monocytes by a TF-kappaB- and HDAC3-dependent mechanism. We conclude that histone deacetylases, particularly HDAC3, play a hitherto unsuspected role in regulating TF expression and raise the possibility that HDACi might be a novel therapy for thrombotic disorders.
组蛋白去乙酰化酶抑制剂（HDACi），如曲古抑菌素A（TSA），可通过促进组蛋白和转录因子的乙酰化来调节基因表达。人组织因子（TF）的表达部分受一个独特的、与核因子-κB相关的“TF-κB”启动子结合位点调控。我们发现TSA和其他四种HDACi（阿皮西丁、MS-275、丁酸钠和丙戊酸）在生理激动剂肿瘤坏死因子（TNF）-α、白细胞介素-1β、脂多糖和HOSCN刺激人脐静脉内皮细胞时，均能抑制约90%的TF活性和蛋白水平诱导，而不影响核因子-κB调节的黏附分子细胞间黏附分子-1（ICAM-1）和E-选择素的表达。TSA和丁酸钠在体外对外周血单核细胞以及在体内对巯基乙酸诱导的小鼠腹腔巨噬细胞中，也能使TF诱导降低约50%。在人脐静脉内皮细胞中，TSA可使TNF-α刺激的TF mRNA转录减弱约70%，而不影响ICAM-1的转录。通过电泳迁移率变动分析，TNF-α和脂多糖诱导强烈的p65/p50和p65/c-Rel异二聚体与核因子-κB和TF-κB探针结合。TSA几乎完全消除TF-κB结合，而不影响核因子-κB结合。染色质免疫沉淀分析和启动子-荧光素酶报告系统证实，TSA抑制TF-κB而非核因子-κB的激活。染色质免疫沉淀和小干扰RNA抑制剂研究表明，HDAC3在TNF-α介导的TF诱导中起重要作用。因此，HDACi通过TF-κB和HDAC3依赖性机制转录抑制激动剂诱导的内皮细胞和单核细胞中TF的表达。我们得出结论，组蛋白去乙酰化酶，尤其是HDAC3，在调节TF表达中发挥了迄今未被怀疑的作用，并提出HDACi可能是血栓性疾病新疗法的可能性。
