Molecular biological studies on environmental chemical pollutants interfering with endocrine systems

我们用分子生物学的方法研究了内分泌干扰物镉。我们的项目包括两项研究：镉暴露诱导的基因表达和慢性镉暴露引起的伊泰-伊泰病的基因分析。[镉暴露诱导的基因表达]为了阐明镉对内分泌系统的影响，我们首先用消减抑制杂交(SSH)的方法筛选了COS-7细胞中对镉反应的基因。我们已经检测到了100多种镉诱导的mRNA物种，并通过测序分析确定了它们的来源。对镉敏感的基因有：金属硫蛋白II、Zeta-晶体、细胞色素C氧化酶亚基IV、谷胱甘肽合成酶、丝氨酸/苏氨酸蛋白激酶、热休克蛋白10、转导β-I亚基、TUNP、Lipocortin II、HSP 10、HSP 40、HSP 60、HSP 86、BAG-3、补体细胞溶解抑制因子(CLI)等。随着镉浓度和暴露时间的变化，它们各自呈现出其特有的表达模式。[伊泰-伊泰病的遗传学分析]为了阐明雌激素受体α在伊泰-伊泰病发病机制中的作用，我们检测了伊泰-伊泰病患者雌激素受体α基因的基因多态性，并与正常对照组进行了比较。我们检测了雌激素受体α基因上游区内含子1的PuvII、XbaI RFLP和AT重复序列的多态性。患者组的基因分布与对照组相似，因此未观察到与Itai病相关的基因分布模式。我们的结论是，该基因的多态性可能与伊泰-伊泰病无关。

We studied an endocrine disruptor, cadmium, by molecular biological methods. Our project included two studies : gene expression induced by cadmium exposure and genetic analyses of Itai-itai disease which is caused by chronic exposure to cadmium.[Gene expression induced by cadmium exposure] To clarify the effects of cadmium on the endocrine system, we firstly screen the genes responsive to cadmium in COS-7 cells with a subtractive suppressive hybridization (SSH) method. We have detected more than 100 mRNA species which were induced by cadmium exposure, and identified their origins by sequencing analysis. The genes responsive to cadmium were as follows : metallothionein II, zeta-crystalline, cytochrome C oxidase subunit IV, glutathione synthetase, serine/threonine protein kinase, heat shock protein 10, transduction beta-I subunit, tunp, lipocortin II hsp 10, hsp 40, hsp 60, hsp 86, BAG-3, complement cytolysis inhibitor (CLI) and so on. Each of them showed its characteristic expression pattern when cadmium concentration and exposure time changed. Further analyses are required to clarify the relationships between the genes responsive to cadmium and the endocrine system.[Genetic analyses of itai-itai disease] In order to clarify the role of estrogen receptor α on the pathogenesis of itai-itai disease, we examined the genotypic polymorphisms in estrogen receptor α gene of the patients with itai-itai disease and compared them with those of control subjects. We examined PuvII, XbaI RFLP polymorphism in intron 1 and AT repeat polymorphism in upstream region of the estrogen receptor α gene. The genotypic distributions of the patient group were similar to those of the control groups, hence no itai-itai disease-related pattern of genotypic distribution was observed. We conclude that polymorphisms of the gene may not be associated with itai-itai disease.