Development of risk assessment system for human by environmental disruptants using nuclear hormone receptor

利用核激素受体开发环境破坏者对人类的风险评估系统

• 批准号: 11839021
• 负责人: DOI Rikuo
• 金额: $ 2.24万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11839021/
• 关键词: endocrine disruptants; thyroid hormone receptor; thyroid hormone response element; T_3; reporter assay; luciferase; hydroxylated PCB; dioxin; 甲状腺ホルモンレセプター; リポーターアッセイ; ホルモンレスポンスエレメント; direct repeat(DR)4; DRO; レチノイン酸Xレセプター; テトラサイクリン

Endocrine disruptants are thought to have various effects on various organs. Estrogenic responses by endocrine disruptants were analyzed using eukaryotic cells and yeast, which are overexpressing human estrogen receptors. In wildlife, some species such as seagulls contaminated with large amount of environmental pollutants showed thyroid hyperplasia and thyroid tumors. To address the effects on thyroid hormone system by endocrine disruptants, the HeLaTR cells that constitutively overexpressed thyroid hormone receptor were constructed and analyzed. Luciferase activity was induced by the addition of T3 in HeLaTR cells transfected with the plasmid containing thyroid hormone response element as an enhancer. Bisphenol A and 2-diethylhexyl phtalate, both could bind to estrogen receptor, had no effect on the luciferase activity, but hydroxylated PCB repressed the luciferase activity caused by the addition of T3. In contrast, dioxin enhanced the luciferase activity, however dioxin alone caused no effect on the activity. These results suggested that HeLaTR cells could be used as the tool for thyroid hormone action by endocrine disruptants.

内分泌干扰物被认为对各种器官有各种影响。雌激素反应的内分泌干扰物进行了分析，使用真核细胞和酵母，这是过度表达人类雌激素受体。在野生动物中，某些物种如海鸥，受到大量环境污染物的污染，表现出甲状腺增生和甲状腺肿瘤。为探讨内分泌干扰物对甲状腺激素系统的影响，构建了组成型过表达甲状腺激素受体的HeLaTR细胞并进行了分析。通过在用含有甲状腺激素反应元件作为增强子的质粒转染的HeLaTR细胞中加入T3来诱导荧光素酶活性。双酚A和邻苯二甲酸二乙基己酯均能与雌激素受体结合，对荧光素酶活性无影响，但羟基化PCB抑制了T3诱导的荧光素酶活性。与此相反，二恶英增强荧光素酶活性，但二恶英单独对活性没有影响。这些结果表明，HeLaTR细胞可用作内分泌干扰物作用甲状腺激素的工具。