剂量敏感的性别反转先天性肾上腺发育不良基因1与雄激素受体相互作用的研究  

Study on interaction between DAX-1 and AR protein

作　　者：杨丽华[1] 李俞池 王旭亮[1] 石敏[1] 江智茂[1] 桂耀庭[1] 

机构地区：[1]北京大学深圳医院男性生殖与遗传重点实验室,深圳518036

出　　处：《中国男科学杂志》2014年第2期7-10,共4页Chinese Journal of Andrology

基　　金：深圳市科创委项目（编号：CXB201104220045A）,深圳市科技计划项目（编号：201202001,201003076）

摘　　要：目的：剂量敏感的性别反转-先天性肾上腺发育不良基因1（dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1, DAX1）和雄激素受体（androgen receptor, AR）在男性生殖系统中均发挥着不可替代的作用，以往研究证明二者之间存在某些联系，本实验主要检测DAX-1和AR蛋白在细胞内的相互作用情况。方法构建人的带HA多肽标签的DAX-1真核表达载体,以脂质体方式将该质粒和雄激素受体真核表达质粒共转染至非洲绿猴肾成纤维细胞（COS-7）中，转染48h后收集细胞提取总蛋白，利用免疫共沉淀技术检测DAX-1和AR蛋白在细胞内的相互作用。结果成功构建了人DAX-1真核表达载体；免疫共沉淀实验结果显示DAX-1和AR蛋白在COS-7细胞内可以相互作用。结论 DAX-1与AR蛋白在细胞内可能存在相互作用，为后续研究DAX-1功能、突变致病及DAX-1与AR的作用机制奠定基础。Objective DAX-1 and AR play irreplaceable roles in male reproductive system, and show some relationship between them. This study is mainly to validate the interaction between DAX-1 and AR protein. Methods DAX-1 eukaryotic expression vector with HA peptide tag were constructed. The recombined plasmid was sequenced and cotransfected into COS-7 cells by lipofectamineTM2000 with the eukaryotic expression vector of AR. After 48 hours, cells were collected and total protein was extracted.Co-Immunoprecipitation technique was used to detect the interaction between DAX-1 and AR protein. Results The eukaryotic expression vector of human DAX1 was constructed successfully. The interaction between DAX-1 and AR was validated by Co-Immunoprecipitation. Conclusion DAX-1 may interact with AR protein. This study might lay the foundation for further study of DAX-1 function, mutation and the molecular mechanism of its interaction with AR.

关 键 词：基因 X连锁 受体 雄激素 男性泌尿生殖系统疾病 

分 类 号：R697[医药卫生—泌尿科学]