Molecular biological studies on environmental chemical pollutants interfering with endocrine systems

环境化学污染物干扰内分泌系统的分子生物学研究

• 批准号: 10670350
• 负责人: NISHIO Hisahide
• 金额: $ 1.66万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670350/
• 关键词: endocline disruptor; cadmium; subtractive suppressive hybridization; itai-itai disease; estrogen receptor; polymorphism; subtractive suppressive hybridization

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.

我们通过分子生物学方法研究了一种内分泌干扰物——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暴露镉引起的痛痛病的遗传分析。[镉暴露诱导的基因表达]为了阐明镉对内分泌系统的影响，我们首先采用消减抑制杂交（SSH）方法筛选COS-7细胞中对镉敏感的基因。我们检测到了100多种因镉暴露而诱导的mRNA种类，并通过测序分析确定了它们的起源。对镉敏感的基因如下：金属硫蛋白 II、zeta 结晶、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 细胞溶解抑制剂（CLI）等。当镉浓度和暴露时间改变时，它们各自表现出其特有的表达模式。需要进一步分析阐明镉敏感基因与内分泌系统之间的关系。 【痛痛病的基因分析】为了阐明雌激素受体α在痛痛病发病机制中的作用，我们检测了痛痛病患者雌激素受体α基因的基因多态性，并与对照受试者进行了比较。我们检测了雌激素受体α基因内含子1的PuvII、XbaI RFLP多态性和上游区域的AT重复多态性。患者组的基因型分布与对照组相似，因此没有观察到与痛痛病相关的基因型分布模式。我们得出结论，该基因的多态性可能与痛痛病无关。

项目成果

Hisahide Nishi et al: "Itai-Itai disease is not associated with polymorphisms of the estrogen receptor α gene"Arch Toxicol. 73. 496-8 (1999)

Hisahide Nishi 等人：“痛痛病与雌激素受体 α 基因的多态性无关”Arch Toxicol. 73. 496-8 (1999)

Hisahide Nishio et al.: "Itai-Itai disease is not associated with polymorphisms of the estrogen receptor α gene"Arch. Toxicol.. 73. 496-498 (1999)

Hisahide Nishio 等人：“痛痛病与雌激素受体 α 基因的多态性无关”Arch. 73. 496-498 (1999)

Hisahide Nishio et al: "Itai-Itai disease is notassociated with polymorphisms of the estrogenreceptor α gene"Arch Toxicol. 73. 496-498 (1999)

Hisahide Nishio 等人：“痛痛病与雌激素受体 α 基因的多态性无关”Arch Toxicol. 73. 496-498 (1999)