基于组蛋白甲基化表观遗传修饰研究芳香烃受体（AhR）介导的致癌分子毒理机制

项目名称： 基于组蛋白甲基化表观遗传修饰研究芳香烃受体（AhR）介导的致癌分子毒理机制

项目编号： No.21277128

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 环境科学、安全科学

项目作者： 靳远祥

作者单位： 浙江工业大学

项目金额： 78万元

中文摘要： 多环芳烃和二噁英通过激活芳香烃受体（AhR）介导的转录途径而诱导疾病如癌症产生。组蛋白去甲基化酶是一些受体的转录共调解因子，但在AhR介导的转录调控及致肿瘤中的作用仍未知。本研究拟通过在细胞中进行基因过表达和抑制表达确认LSD1和Jmjd1a对AhR转录活性影响；用免疫沉淀技术论证LSD1或Jmjd1a在细胞内和AhR的蛋白质互作效应；进一步用染色质免疫沉淀技术分析当AhR转录通路激活时，其靶标基因增强子和启动子区域组蛋白甲基化修饰水平的变化，并通过基因抑制LSD1和Jmjd1a表达确定它们在AhR靶标基因增强子或启动子区域去甲基化的修饰位点和状态。用动物实验进一步论证LSD1和Jmjd1a在配体激活的AhR转录通路中的作用并解析其在肿瘤发生的作用。通过该项目的实施明确组蛋白去甲基化酶在AhR介导的基因转录调控和信号转导中的功能，为解析由多环芳烃等诱导基因异常表达和癌症发生机制提供证据。

中文关键词： 多环芳烃受体；LSD1；Jmjd1a；组蛋白修饰；基因表达调控

英文摘要： Aryl hydrocarbon receptor (AhR) is a ligand-dependent transcription factor that plays important role in normal physiology with endogenous ligands and mediates the adverse effects of several classes of exogenous ligands including PAHs and dioxins. Transcriptional co-regulators mediate gene expression through covalent histone modifications. However, involvement of histone methyltransfeases (HMTs) and demethylases (HDMs) in AhR-mediated transcription still remains unknown. Firstly, to screen co-regulator activity of HMTs/HDMs for AhR-mediated transcription, the expression vector of different HDMs including LSD1, Jmjd1a, Jmjd1b, Jmjd1c, Jmjd2b, Jmjd2c, Jmjd2d, Jarid1a, Jarid1b, Jarid1c, Jarid1d and HMTs including Mll5 or Suv39H1 together with AhR reporter vector were co-transfected into 293F cells and analyzed by Luciferase reporter assay, respectively. According to the results, we observe that LSD1 and Jmjd1a promotes the transcriptional activity of AhR as a co-activator without ligand specific. In the present study, the following plans will be adopted to elucidate the molecular mechanism of histone demethy lation mediate the transcription of AhR in cells and mouse.1) To understand whether LSD1 or Jmjd1a is able to interact with AhR in cellular environments, immunoprecipitation (IP) will be performed after transien

英文关键词： Aryl hydrocarbon receptor；LSD1；Jmjd1a；histone modifications；gene transcription