Development of New Screening Method for Endocrine Disruptors and Analyses of Disrupting Mechanisms

内分泌系统参与多种生理过程。近年来，内分泌干扰物被认为在这些过程中发挥了一定的作用。因此，重要的是开发具有这些活性的化学品的筛选方法，并且还表征干扰机制。1）我们使用酵母双杂交系统开发了具有激素活性的化学品的新筛选方法。2）使用该测定系统，我们对500多种化合物进行了结构-活性关系的研究过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptor γ）是核受体之一，具有双重DNA结合特异性。当过氧化物酶体增殖物激活受体γ使同源二聚体结合雌激素受体结合序列而不是直接重复。利用该细胞系，我们采用消减法和双向电泳技术分离了新的PPARγ靶基因。

The endocrine systems are involved in the multitude of physiological processes. Recently, the endocrine disrupting chemicals are thought to have some effects in these processes. Therefore, it is important to develop a screening method for chemicals having these activities, and also characterize a disrupting mechanism.1) We developed a new screening method for chemicals having hormonal activities using yeast two-hybrid system.2) Using this assay system, we tested more than five hundred chemicals and discussed the structure-activity relationship.3) We demonstrated that PPAR_γ (peroxisome proliferator-activated receptor γ), which is one of the nuclesr receptor, has dual DNA binding specificity. When PPARγ makes homodimer binds to estrogen receptor binding sequences rather than direct repeat.4) We established PPARγ-expressing cells. Using this cell line, we isolated the new target genes of PPARγ using subtraction protocol and two-dimensional electrophoresis.