中国人21-羟化酶缺乏症基因型和临床表型特点研究  

Molecular Characterization of Mutations and Phenotype/Genotype Correlation in Chinese Patients with 21-hydroxylase Deficiency

作　　者：张波[1] 陆召麟[1] 王玥[1] 陶红[1] 

机构地区：[1]中国医学科学院中国协和医科大学北京协和医院内分泌科,北京100730

出　　处：《Acta Genetica Sinica》2004年第9期950-955,共6页

摘　　要：利用快速可靠的基因突变检测方法研究中国人 2 1 羟化酶缺乏症 (2 1 hydroxylasedeficiency ,2 1 OHD)基因突变特点及基因型和临床表型的关系。用 8例非经典型患者、35例经典型患者及 34例正常对照者基因组DNA作模板 ,用特定引物特异性扩增CYP2 1的两个片段 ,片段 1从Exon1→Exon3,片段 2从Exon3→Exon10。用片段 1和片段 2为模板进行第二轮PCR ,用特定限制性内切酶消化后经琼脂糖凝胶电泳鉴定 9种突变 ,包括Del、Exon3Del8bp、Q318X、R35 6W、Exon6Cluster、i2g、I172N、P30L和V2 81L。结果表明 ,4 3例患者的 86个等位基因中的 79个(91 9% )检测出突变 ,最常见的是I172N(36 0 % ) ,其次为i2g(2 0 9% )、Del(8 6 % )、P30L(7 0 % )、Q318X(7 0 % )、V2 81L(4 7% )、R35 6W(2 3% )、E6Cluster(2 3% )和Exon3Del8bp(1 2 % )。失盐型、单纯男性化型和非经典型 2 1 羟化酶缺乏症各临床分型中最常见的突变分别是Del(4 4 4 % )、I172N(4 4 2 % )和P30L(37 5 % )。另外根据对 2 1 羟化酶活性影响程度将基因型分为轻、中、重 3组 ,3组间初诊年龄、17 羟孕酮 (17 OHP)、该病亚型构成均存在显著差异 (P <0 .0 5 ) ,提示基因型决定临床表型。研究结果表明 ,中国人 2 1 OHD最常见的突变为I172N、i2g和Del,中?To investigate characteristics of CYP21 mutations in Chinese patients with 21-hydroxylase deficiency(21-OHD) and the relationship between genotype and phenotype,a time saving,reliable molecular diagnosis method was developed.Eight patients with NC 21-OHD,35 patients with classical ones and 34 normal controls were studied as followed: CYP21 gene was amplified into fragment 1 (exon1→exon3) and fragment 2 (exon3→exon10) through PCR with specific primers.The second round PCR was carried out using fragment 1 and 2 as template,and PCR products were digested by restrictive enzymes to analyze nine mutations,including Del,Exon3 Del8 bp,Q318X,R356W,Exon6Cluster,i2g,I172N,P30L and V281L.The most frequent mutation in 43 cases of Chinese 21-OHD was I172N(36.0%),followed by i2g(20.9%), Del(8.6%),P30L(7.0%),Q318X(7.0%),V281L(4.7%),R356W(2.3%),E6Cluster(2.3%)and Exon3 Del8 bp(1.2%).The most common CYP21 gene defect in salt wasting,simple virilizing and nonclassical form was Del(44.4%),I172N(44.2%) and P30L(37.5%)respectively.In addition,genotype was classified into three (severe,moderate and mild) groups according to the degree of 21-hydroxylase enzymatic compromise caused by the mutation.Age of first visit,basal 17-OHP,composition of disease subtype were significantly different among the three groups, P <0.05.In conclusion,the most common mutations in Chinese 21-OHD were I172N,i2g and Del.The genotype of Chinese 21-OHD has strong correlation with the phenotype.

关 键 词：21—羟化酶缺乏症 先天性肾上腺皮质增生症 甾类21—单加氧酶 CYP21基因 

分 类 号：R586[医药卫生—内分泌]