FANCL新突变(c.1033G>A)致早发性卵巢功能不全1例报道及体外功能验证  

A case report of premature ovarian insufficiency caused by a novel FANCL mutation(c.1033G>A)and in vitro functional validation

作　　者：刘怡箐 任淑婷 潘韵程 张锋 张晓金 吴燕华[1,3] LIU Yi-qing;REN Shu-ting;PAN Yun-cheng;ZHANG Feng;ZHANG Xiao-jin;WU Yan-hua(Department of Genetics,Fudan University,Shanghai 200433,China;Department of Gynecology,Obstetrics and Gynecology Hospital,Fudan University,Shanghai 200011,China;National Demonstration Center for Experimental Biology Education,School of Life Sciences,Fudan University,Shanghai 200433,China)

机构地区：[1]复旦大学生命科学学院遗传学系,上海200433 [2]复旦大学附属妇产科医院妇科,上海200011 [3]复旦大学生物科学国家级实验教学示范中心,上海200433

出　　处：《复旦学报(医学版)》2025年第2期270-276,291,共8页Fudan University Journal of Medical Sciences

基　　金：国家重点研发计划(2022YFC2703800);国家自然科学基金(32270658)。

摘　　要：目的探究一例早发性卵巢功能不全(premature ovarian insufficiency,POI)患者中发现的FANCL新突变的特点,并在体外研究其对功能的影响。方法利用全外显子组测序(whole-exome sequencing,WES)技术,在一例POI患者中筛选到了新的FANCL杂合突变c.1033G>A(p.Glu345Lys),家系回访发现该突变遗传自早绝经的母亲。通过sanger测序鉴定该突变真实性,并通过软件预测突变位点的保守性。构建过表达FANCL突变型和野生型质粒,瞬时转染进HEK293T细胞株,通过qPCR、免疫荧光和Western blot来检测突变产生的影响。结果该突变位于FANCL的环状结构域内且在多物种中高度保守。突变体mRNA表达水平没有明显变化,而蛋白质表达水平显著下调。体外细胞实验进一步揭示该变异会通过降低蛋白质稳定性导致表达水平下降。结论该POI患者存在FANCL c.1033G>A变异,并引起蛋白质稳定性下降而导致患者患病。Objective To investigate the characteristics of a novel FANCL mutation identified in a patient with premature ovarian insufficiency(POI)and to explore its potential functional impacts in vitro.Methods A novel FANCL heterozygous mutation c.1033G>A(p.Glu345Lys)was screened in a patient with POI using whole exome sequencing(WES),which was found to be inherited from a mother who had undergone early menopause.The authenticity of the mutation was identified by Sanger sequencing and the conserved nature of the mutation site was predicted by software.Overexpressing FANCL mutant and wildtype plasmids were constructed and transiently transfected into HEK293T cell lines,and the effect of the mutation was detected by qPCR,immunofluorescence and Western blot.Results The mutation site of FANCL was located within the Ring domain of FANCL,which was highly conserved across multiple species.The mutant showed no significant change in mRNA expression level,while the protein expression level was significantly down-regulated.In vitro cellular experiments further revealed that the mutation leads to decreased expression levels by reducing protein stability.Conclusion A FANCL c.1033G>A mutation was found and it may cause disease in the POI patient due to decreased protein stability.

关 键 词：早发性卵巢功能不全(POI) FANCL 全外显子组测序(WES) 错义突变 蛋白质稳定性 

分 类 号：R394[医药卫生—医学遗传学]