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DNA tip analysis for the polymorphisms of osteoporosis

骨质疏松症多态性的 DNA 尖端分析

基本信息

批准号：
11557202
负责人：
MIYAMOTO Ken-ichi
金额：
$ 8.7万
依托单位：
Tokushima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557202/
关键词：
vitamin D receptor DNA analysis osteoporosis polymorphism calcium absorption DNA マイクロチップ FRET

项目摘要

To analyze the gene polymorphism for osteroprosis, we developed FRET based polymerase chain reaction for multiple region of the polymorphism. In addition, we identified the polymorphism of the human vitamin D receptor gene promoter. The major physiological activity of 1,25-hydroxyvitamin D3 (1,25(OH)2D3) is the regulation of calcium absorption in the small intestine, and the level of vitamin D receptor (VDR) is an important factor in this regulation. We demonstrated that the caudal-related homeodomain Cdx-2 played an important role in the intestine-specific transcription of the human VDR gene. In the present project, the polymorphism was identified in the core sequence in the Cdx-2 binding site in the VDR gene promoter. In 261 Japanese women with genotyped VDR polymorphisms, 48 were genotype Cdx-A (adenine at -3731 nt relative to the transcription start site of human VDR gene), 82 were genotype Cdx-G (guanine at -3731 nt), 131 were genotype Cdx-A/G (heterozygote). In postmenopausal Japanese women, the bone mineral density (BMD) in the lumbar spine (L2-4) with the Cdx-G homozygote was 12% lower than that with the Cdx-A homozygote (P<0.05). In electrophoretic gel mobility shift assay, the oligonucleotide with Cdx-G allele markedly decreased the binding to Cdx-2 compared with that in the Cdx-A allele. The transcriptional activity of the VDR promoter with Cdx-G allele was decreased to 70% of the Cdx-A allele. In addition, in the herpes simplex virus thymidine kinase promoter, the Cdx-2 binding element with the G allele showed significantly lower transcriptional activity than that of the A allele. Thus, the polymorphism in the Cdx-2 binding site of the VDR gene (Cdx-polymorphism) would affect the expression of VDR in the small intestine. In addition, this polymorphism may modulate BMD in postmenopausal Japanese women.

为了分析骨质疏松症的基因多态性，我们开发了基于FRET的聚合酶链反应的多个区域的多态性。此外，我们确定了人类维生素D受体基因启动子的多态性。1，25-羟基维生素D3（1，25（OH）2D 3）的主要生理活性是调节小肠内钙的吸收，维生素D受体（VDR）水平是调节小肠内钙吸收的重要因素。我们证明，尾相关的同源结构域Cdx-2在人类VDR基因的精氨酸特异性转录中发挥了重要作用。在本项目中，在VDR基因启动子Cdx-2结合位点的核心序列中鉴定了多态性。在261例VDR基因多态性的日本女性中，48例为Cdx-A（人类VDR基因转录起始位点-3731 nt处的腺嘌呤）基因型，82例为Cdx-G（-3731 nt处的鸟嘌呤）基因型，131例为Cdx-A/G（杂合子）基因型。在绝经后的日本妇女中，Cdx-G纯合子的腰椎（L2-4）骨密度（BMD）比Cdx-A纯合子低12%（P<0.05）。凝胶电泳迁移率变动分析表明，与Cdx-A等位基因相比，Cdx-G等位基因的寡核苷酸与Cdx-2的结合能力明显降低。Cdx-G等位基因的VDR启动子的转录活性降低至Cdx-A等位基因的70%。此外，在单纯疱疹病毒胸苷激酶启动子中，Cdx-2结合元件与G等位基因的转录活性显着低于A等位基因。因此，VDR基因Cdx-2结合位点的多态性（Cdx-多态性）将影响小肠中VDR的表达。此外，这种多态性可能调节绝经后日本妇女的BMD。