在性早熟女孩中检测到一种新的雌激素受体基因突变。

Bing Li, Li Liu, Xin Fu, Wen-Qu Zhou, Dong-Ting Zou, Xiao-Yuan Zhao, Yan-Na Cai, Hong-Bin Tu, Qi-Cai Liu, Yao-Yong Chen
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引用次数: 0

摘要

女性性早熟是由于下丘脑-垂体-性腺轴过早激活,暴露于外源性性类固醇激素,以及各种因素引起的内源性性类固醇的存在而引起的。雌激素是青春期开始的最后一个关键因素。然而,在一些女孩性早熟的病例中,雌激素升高无法检测到。雌激素受体敏感性升高,可能由ESR1突变或多态性引起,已被频繁提及用于解释散发性低雌激素型病例的病因。但在临床实践中未发现病例证据。为了筛选雌激素受体基因可能的突变,收集16例散发性低雌激素性早熟女孩的白细胞基因组DNA,扩增ESR1外显子,采用PCR-SSCP/银染色法进行分析。在其中一名患者(No. 14)中发现外显子8的单链构象改变。将该可疑片段克隆到T载体上并进行测序分析。对这些克隆的测序显示,这种构象变化是由C到T的转变引起的。这种突变导致ESR1蛋白548位的精氨酸被胱氨酸取代。该突变产生了一个额外的Btsl消化位点,使其易于用PCR-PFLP方法鉴定。利用该方法进一步检测,并对患者克隆的外显子8菌落进行测序,证实14号患者基因型为Arg548/Cys548异源型。这种突变极大地增加了该地区的疏水性。该残基在脊椎动物中的位置和保守性表明Arg548可能在ESR1功能中起重要作用。为了研究该突变在性早熟发病中的作用,我们构建了萤火虫荧光素酶报告质粒pgl3启动子- ere,以及pCR3。以野生型pCR3为基础构建表达cys548er的1-hermut pisimid。1她。报告基因与pCR3的共转染。1 -hermut在CMF-7细胞株中的表达证明,Cys548突变体比Arg548野生型的转录活性显著提高。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A novel mutation of estrogen receptor gene detected in girls with precocious puberty.

Female precocious puberty is caused by premature activation of the hypothalamic-pituitary-gonadal axis, exposure to exogenous sex steroid hormones, and the presence of endogenous sex steroids caused by various factors. Estrogen is the final key factor to start onset of puberty. However,in some cases of precocious puberty in girls estrogen elevation could not be detected. The raised sensitivity of estrogen receptor, which may caused by ESR1 mutation or polymorphism, has been frequently mentioned for interpreting the etiology of sporadic low estrogen type cases. But no case evidence has been found in clinical practice. For the purpose of screening possible mutations in estrogen receptor gene, leukocyte genomic DNA were collected from 16 girls with precocious puberty of sporadic low estrogen,and exons of ESR1 were amplified and analysized using PCR-SSCP/silver staining method. A single strand conformation change in exon 8 was found in one of the patients (No. 14). The suspected fragment were cloned to a T vector and sequenced for analysis. Sequencing of these clones revealed that this conformation change is caused by a C to T transition. This mutation results in the replacement of arginine by cystine at position 548 of ESR1 protein. The mutation created an extra Btsl digest site and made it can be readily identified by PCR-PFLP method. Further detection using this method, and sequencing of cloned exon8 colonies from patients proved that the patient No. 14 is Arg548/Cys548 heterozagous in genotype. This mutation increased hydrophobility of the area dramatically. The position and the conservative of this residue in vertebrates suggested Arg548 may play an important role in ESR1 function. For study the role of this mutation in the onset of precocious puberty, a firefly luciferase reporter plasmid pGL3-promoter-ERE was constructed,and a pCR3. 1-hermut pisimid expressing Cys548 ER was constructed based on wild type pCR3. 1her. Co-transfection of reporter and pCR3. 1 -hermut in CMF-7 cell strain proved that Cys548 mutant can significantly increase the transcription activity over the Arg548 wild type.

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