中国南方汉族激素耐药型肾病综合征儿童WT1基因突变分析

发布时间：2018-06-02 09:25

本文选题：激素耐药型肾病综合征 + 单卵双胞胎　；参考：《福建医科大学》2012年硕士论文

【摘要】：第一部分激素耐药型肾病综合征单卵孪生姐妹WT1基因突变分析目的分析1个南方汉族激素耐药型肾病综合征（SRNS）家系WT1基因突变及其特点。方法研究对象为1个南方汉族SRNS家系先证者及其单卵孪生姐姐、父母和大姐，对照人群为50例尿检正常的汉族成年人。取所有研究对象外周静脉血3mL，提取基因组DNA，PCR扩增WT1基因全部10个外显子及其周围的部分内含子序列，对PCR产物直接进行DNA序列测定。结果先证者临床表型为不完全型Denys-Drash综合征，其孪生姐姐临床表型为孤立性SRNS。在该对单卵孪生姐妹均检测出WT1基因1180CT（R394W）杂合突变，在先证者父母及其大姐和50例对照人群未检测出1180CT（R394W）杂合突变；此外，还在该对孪生姐妹检测出3个相同的WT1基因多态性—126CT、903AG和IVS7 32CA。结论本研究在1个汉族SRNS家系临床表型不同的单卵孪生姐妹患儿发现了相同的WT1基因R394W杂合突变。第二部分中国汉族散发性孤立性激素耐药型肾病综合征女性患儿WT1基因突变分析目的分析14例汉族散发性孤立性激素耐药型肾病综合征（ISRNS）女性患儿WT1基因突变及其特点。方法研究对象为14例汉族散发性ISRNS女性患儿（已除外NPHS2基因突变），对照人群为38例汉族散发性ISRNS男孩和50例尿检正常的汉族成年人。取所有研究对象外周静脉血3mL，提取基因组DNA，PCR扩增WT1基因外显子8和9及其周围的部分内含子序列，对PCR产物直接进行DNA序列测定。结果对14例汉族散发性ISRNS女性患儿进行WT1基因突变检测，结果在1例散发性ISRNS女孩患儿检测出WT1基因IVS9+5GA杂合变异，该变异在50例正常对照人群中未检测出。汉族散发性ISRNS女性患儿WT1基因突变检出率7.1%。结论中国汉族散发性ISRNS女性患儿存在WT1基因突变，提示对汉族散发性ISRNS女性患儿需进行WT1基因突变分析。第三部分1例汉族孤立性肾病综合征伴肾衰竭男性患儿WT1基因突变分析目的分析1例汉族孤立性肾病综合征伴肾衰竭男性患儿WT1基因突变及其特点。方法研究对象为1例汉族孤立性肾病综合征伴肾衰竭男性患儿，6.3岁发病，，对照人群为110例尿检正常的汉族成年人。取所有研究对象外周静脉血3mL，提取基因组DNA，PCR扩增WT1基因全部10个外显子及其周围的部分内含子序列，对PCR产物直接进行DNA序列测定。结果在该患儿WT1基因外显子8检测出1051AG杂合变异，但在110例正常对照人群中未检测出1051AG变异，表明该变异为WT1基因突变。该突变导致WT1基因的编码蛋白WT1的第351位赖氨酸被谷氨酸取代，即K351E。经检索PUBMED文献数据库、单核苷酸多态性数据库和人类基因突变数据库已公布的WT1基因变异，证实1051AG（K351E）为新发现的WT1基因突变。结论在1例6.3岁孤立性肾病综合征伴肾衰竭男性患儿检测出WT1基因突变，提示对孤立性肾病综合征伴肾衰竭男性患儿需进行WT1基因突变分析。
[Abstract]:Analysis of WT1 gene mutation in monozygotic twin sisters of steroid-resistant nephrotic syndrome Objective to analyze the mutation and characteristics of WT1 gene in a Chinese family with steroid-resistant nephrotic syndrome. Methods the subjects of this study were a proband from a SRNS family of southern Han nationality and their monozygotic twin sisters, parents and elder sisters, and a control group of 50 Han adults with normal urine tests. All 10 exons of WT1 gene and some intron sequences around them were amplified by genomic DNA-polymerase chain reaction (DNA-PCR) from peripheral venous blood of all subjects. The PCR products were sequenced by DNA directly. Results the clinical phenotype of the proband was incomplete Denys-Drash syndrome, and the twin sister's clinical phenotype was isolated. Heterozygous mutations of WT1 gene were detected in all of the single twin sisters, and no heterozygous mutations were detected in the proband parents and their elder sisters and 50 controls. Three identical WT1 gene polymorphisms-126 CTG 903AG and IVS7 32 CA were also detected in the twin sisters. Conclusion the same R394W heterozygous mutation of WT1 gene was found in a single twin sister with different phenotypic phenotypes in a SRNS pedigree of Han nationality. The second part of the Chinese Han nationality sporadic isolated hormone resistant nephrotic syndrome in women with WT1 gene mutation analysis Objective to analyze the mutation and characteristics of WT1 gene in 14 children with sporadic isolated steroid-resistant nephrotic syndrome (ISR NSS) of Han nationality. Methods the subjects were 14 sporadic ISRNS female children (excluding NPHS2 gene mutation), 38 Han sporadic ISRNS boys and 50 normal controls. The exons 8 and 9 of WT1 gene and some intron sequences around them were amplified by PCR from peripheral venous blood of all subjects. The PCR products were sequenced by DNA directly. Results the mutation of WT1 gene was detected in 14 sporadic ISRNS female children of Han nationality. The results showed that the IVS9 5GA heterozygosity of WT1 gene was detected in 1 sporadic ISRNS girl, but not in 50 normal controls. The detection rate of WT1 gene mutation in sporadic ISRNS women of Han nationality was 7.1%. Conclusion there is a mutation of WT1 gene in sporadic ISRNS female children of Han nationality in China, suggesting that WT1 gene mutation analysis should be carried out in sporadic ISRNS female children of Han nationality. Part three: analysis of WT1 gene mutation in children with isolated nephrotic syndrome with renal failure in Han nationality Objective to analyze the WT1 gene mutation and its characteristics in a patient with isolated nephrotic syndrome with renal failure in Han nationality. Methods one patient with isolated nephrotic syndrome associated with renal failure in Han nationality aged 6.3 years was studied, and 110 Han adults with normal urine test were enrolled in the study. All 10 exons of WT1 gene and some intron sequences around them were amplified by genomic DNA-polymerase chain reaction (DNA-PCR) from peripheral venous blood of all subjects. The PCR products were sequenced by DNA directly. Results 1051AG heterozygosity was detected in exon 8 of WT1 gene, but no 1051AG mutation was detected in 110 normal controls, indicating that the mutation was a mutation of WT1 gene. The mutation resulted in the substitution of lysine at position 351 of WT1, the encoding protein of WT1 gene, by glutamate, K351E. By searching PUBMED literature database, single nucleotide polymorphism database and human gene mutation database, it was confirmed that 1051 AGN K351E was a new WT1 gene mutation. Conclusion WT1 gene mutation was detected in a 6.3 year old boy with solitary nephrotic syndrome with renal failure, suggesting that WT1 gene mutation should be analyzed in the isolated nephrotic syndrome with renal failure.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R726.9

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