1例自身免疫性多内分泌腺病综合征Ⅰ型家系的AIRE基因突变分析  

Analysis of AIRE gene mutation in a pedigree with autoimmune polyendocrine syndrome type Ⅰ

作　　者：黄朱亮 李杨[2] 郑芳[3] HUANG Zhuliang;LI Yang;ZHENG Fang(Department of Clinical Laboratory,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350005,Fujian;Department of Endocrine,Zhongnan Hospital,Wuhan University,Wuhan 430071,Hubei,China;Center for Gene Diagnosis & Department of Clinical Laboratory,Zhongnan Hospital,Wuhan University,Wuhan 430071,Hubei,China)

机构地区：[1]福建医科大学附属第一医院检验科,福州350005 [2]武汉大学中南医院内分泌科,武汉430071 [3]武汉大学中南医院临床基因诊断中心&检验科,武汉430071

出　　处：《临床检验杂志》2019年第8期612-616,共5页Chinese Journal of Clinical Laboratory Science

基　　金：武汉大学中南医院科技创新培育基金(cxpy20160044)

摘　　要：目的分析1例自身免疫性多内分泌腺病综合征Ⅰ型家系的自身免疫调节因子(autoimmune regulator,AIRE)基因突变。方法采集家系成员外周血标本,PCR扩增和Sanger测序筛查AIRE上的突变位点,采用构建酶切位点PCR(created restriction site PCR,CRS-PCR)和PCR-限制性片段长度多态性法(PCR restriction fragment length polymorphism,PCR-RFLP)在100例健康人群中筛查验证,应用SIFT、PolyPhen-2、Mutation Taster、Antheprot Editor软件分析突变对蛋白质结构和功能的影响,使用Alternative Splice Site Predictor、FruitFly Splice Predictor、SplicePort软件预测突变对mRNA剪切位点的影响,并以Sanger测序验证。结果在先证者AIRE基因上筛选出新的复合性杂合突变c.47 C>G T16R和c.1631-2 A>T。c.47位点在不同物种中高度保守和同源性,c.47C>G的错义突变改变蛋白质的二级结构和疏水性,影响蛋白质的功能。c.1631-2 A>T突变会引起剪切位点消失,造成靶mRNA上13号外显子缺失。结论在自身免疫性多分泌腺病综合征Ⅰ型的家系中发现2个新的致病突变位点,分别为c.47C>G T16R和c.1631-2 A>T。Objective To analyze the mutation of autoimmune regulator ( AIRE ) gene in a pedigree with autoimmune polyendocrine syndrome type Ⅰ(APS-Ⅰ). Methods The peripheral blood samples from family members were collected for DNA extraction, and then the mutation sites on AIRE gene were screened by PCR and Sanger sequencing. The mutation sites were further verified in 100 healthy persons by the created restriction site PCR (CRS-PCR) and PCR restriction fragment length polymorphism (PCR-RFLP). The effects of mutation on the structure and function of AIRE protein were analyzed with SIFT, PolyPhen-2, Mutation Taster and Antheprot Editor softwares. The effects of mutation on the splicing sites of AIRE mRNA were predicted with Alternative Splice Site Predictor, FruitFly Splice Predictor and SplicePort softwares, and further verified by Sanger sequencing. Results Two novel heterozygous mutations c.47 C>G T16R and c.1631-2 A>T were found in the proband. The c.47 site is highly conserved and homologous in different species. The missense mutation of c.47C>G changed the secondary structure and hydrophobicity of AIRE protein, and affected its function. The c.1631 -2 A>T mutation changed the splicing site of AIRE mRNA, and led to the deletion of exon 13. Conclusion Two novel pathogenic mutations c.47 C>G T16R and c.1631-2 A>T are identified in a pedigree with autoimmune polyendocrine syndrome type Ⅰ.

关 键 词：自身免疫性多内分泌腺病综合征Ⅰ型 基因诊断 自身免疫调节因子 

分 类 号：R446.9[医药卫生—诊断学]