Molecular biological studies on environmental chemical pollutants interfering with endocrine systems

我们用分子生物学方法研究了内分泌干扰物镉。我们的研究包括两个项目：镉暴露诱导的基因表达和慢性镉暴露引起的Itai-itai病的遗传分析。[Gene为了阐明镉对内分泌系统的影响，我们首先利用消减抑制杂交（SSH）技术筛选了COS-7细胞中对镉敏感的基因。我们检测了100多种镉暴露诱导的mRNA，并通过测序分析确定了它们的来源。对镉有反应的基因如下：金属硫蛋白II、ζ-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、脂皮质素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，它们均表现出其特有的表达模式。需要进一步的分析，以澄清镉和内分泌系统的基因之间的关系。【手足病的遗传学分析】为了阐明雌激素受体α在手足病发病中的作用，我们检测了手足病患者雌激素受体α基因的基因型多态性，并与对照组进行了比较。检测了雌激素受体α基因内含子1的Puv Ⅱ、Xba Ⅰ RFLP多态性和上游AT重复序列多态性。患者组的基因型分布与对照组相似，因此未观察到与itai-itai病相关的基因型分布模式。我们的结论是，基因多态性可能与itai-itai病。

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.