SOX10基因c.661_664dup (p.P222Lfs*60)新变异致病机制的探索

Q4 Medicine
Huiying Li, Peipei Chen, Pingping Liu, Shanshan Yu, Xiaodan Jin, Shuang Zhao
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引用次数: 0

摘要

目的:通过体外实验,探讨SOX10基因c.661_664dup (p.P222Lfs*60)变异体致4C型Waardenburg综合征患儿的发病机制。方法:选取邯郸市第一医院诊断的1例患儿作为研究对象。收集患儿的临床资料。采集了患儿及其父母的外周血样本。提取基因组DNA后,进行三全外显子组测序。候选变异的致病性参照美国医学遗传与基因组学学会(ACMG)指南,通过生物信息学分析确定。候选变异通过Sanger测序进行验证。构建SOX10野生型和c.661_664dup (p.p 2222lfs *60)变异体表达质粒,瞬时转染293T细胞,检测其RNA和蛋白水平的表达。将瞬时转染野生型/突变型SOX10的293T细胞分别用10 ug/mL的环己亚胺(CHX)处理0、4、8、24 h,用Western blotting法检测靶蛋白的降解率。本研究已获邯郸市第一医院伦理委员会批准(伦理号::hdyy - lw - 25053)。结果:患儿携带一种SOX10基因杂合c.661_664dup (p.P222Lfs*60)变异,此前未见报道。该变异在mRNA水平上没有显著改变SOX10的表达,但在蛋白水平上有显著改变。CHX处理后,突变体SOX10蛋白的降解速度减慢。结论:突变体SOX10可能通过影响其蛋白产物的降解速率来影响下游基因的表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Exploration of the pathogenic mechanism of a novel c.661_664dup (p.P222Lfs*60) variant of SOX10 gene].

Objective: To explore the pathogenic mechanism of a child with Waardenburg syndrome type 4C due to a c.661_664dup (p.P222Lfs*60) variant of SOX10 gene through in vitro experiments.

Methods: A child diagnosed at the Handan First Hospital was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples were collected from the child and his parents. Following extraction of genomic DNA, trio-whole exome sequencing was carried out. Pathogenicity of candidate variant was determined by bioinformatic analysis and reference to the guidelines from the American College of Medical Genetics and Genomics (ACMG). Candidate variant was verified by Sanger sequencing. Expression plasmids of wild-type SOX10 and the c.661_664dup (p.P222Lfs*60) variant were constructed and transiently transfected into 293T cells to determine the expression at the RNA and protein levels. The 293T cells transiently transfected with the wild-type/mutant SOX10 were treated with 10 ug/mL cycloheximide (CHX) for 0, 4, 8, 24 h, respectively, and the degradation rate of target protein was detected by Western blotting assay. This study has been approved by the Ethics Committee of Handan First Hospital (Ethics No. HDYY-LW-25053).

Results: The child was found to harbor a heterozygous c.661_664dup (p.P222Lfs*60) variant of the SOX10 gene, which was unreported previously. The variant did not significantly alter the expression of SOX10 at the mRNA level but the protein level. After the CHX treatment, the degradation of mutant SOX10 protein had slowed down.

Conclusion: The mutant SOX10 may affect the expression of downstream genes by affecting the degradation rate of its protein product.

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来源期刊
中华医学遗传学杂志
中华医学遗传学杂志 Medicine-Medicine (all)
CiteScore
0.50
自引率
0.00%
发文量
9521
期刊介绍: Chinese Journal of Medical Genetics is a medical journal, founded in 1984, under the supervision of the China Association for Science and Technology, sponsored by the Chinese Medical Association (hosted by Sichuan University), and is now a monthly magazine, which attaches importance to academic orientation, adheres to the scientific, scholarly, advanced, and innovative, and has a certain degree of influence in the industry. Chinese Journal of Medical Genetics is a journal of Peking University, and is now included in Peking University Journal (Chinese Journal of Humanities and Social Sciences), CSCD Source Journals of Chinese Science Citation Database (with extended version), Statistical Source Journals (China Science and Technology Dissertation Outstanding Journals), Zhi.com (in Chinese), Wipu (in Chinese), Wanfang (in Chinese), CA Chemical Abstracts (U.S.), JST (Japan Science and Technology Science and Technology), and JST (Japan Science and Technology Science and Technology Research Center). ), JST (Japan Science and Technology Agency), Pж (AJ) Abstracts Journal (Russia), Copernicus Index (Poland), Cambridge Scientific Abstracts, Abstracts and Citation Database, Abstracts Magazine, Medical Abstracts, and so on.
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