先天性巨指(趾)症全外显子组测序分析  

Whole exome sequencing analysis of congenital macrodactyly

作　　者：李建峰[1] 田光磊[2] 李大村[1] 潘慧[3] 张文桐[1] 刘井达[1] 赵亮[1] 李冬梅[1] Li Jianfeng;Tian Guanglei;Li Dacun;Pan Hui;Zhang Wentong;Liu Jingda;Zhao Liang;Li Dongmei(Department of Hand Surgery,Beijing Shunyi District Hospital,Beijing 101300,China;Department of Hand Surgery,Beijing Jishuitan Hospital,Beijing 100035,China;Department of Pathology y Beijing Shunyi District Hospital,Beijing 101300,China)

机构地区：[1]北京市顺义区医院手外科,北京101300 [2]北京积水潭医院手外科,北京100035 [3]北京市顺义区医院病理科,北京101300

出　　处：《中华手外科杂志》2022年第1期71-76,共6页Chinese Journal of Hand Surgery

基　　金：北京市顺义区科委科技三项费项目(KS201935)。

摘　　要：目的利用全外显子测序技术研究先天性巨指(趾)症的突变基因和位点。方法将2018年6月至2020年5月在我院手术的12例先天性巨指(趾)症患者分为单纯巨指、单纯巨趾、巨指并指、巨趾并趾4组,对患者的病变组织和外周血进行全外显子检测,将4组检测到的突变基因进行交集,对交集结果进行Phenolyzer分析,筛选巨指(趾)症的致病基因,并对外显子测序结果进一步行Sanger测序验证,明确先天性巨指(趾)症的突变基因和位点,对突变基因所编码的蛋白信号通路进行单克隆抗体免疫组化分析,以多指患儿脂肪细胞作为对照组进行比较,观察目标蛋白表达情况。结果根据全外显子测序综合生物信息分析及Sanger验证结果,筛选出PIK3CA基因突变为巨指(趾)症的致病基因,突变位点为10号外显子c.G1624A(p.E542K)、c.G1633A(p.E545K);21号外显子c.A3140G(p.H1047R)、c.G3145C(p.G1049R),其中c.G3145C(p.G1049R)位点是首次发现,病变组织的AKT单克隆抗体免疫组化分析显示巨指的AKT蛋白表达较多指明显增强。结论PIK3CA基因突变所导致的PI3k-AKT信号通路过度表达是导致先天性巨指(趾)症的原因。Objective To study the mutated genes and loci of congenital macrodactyly by whole exon sequencing technique.Methods From June 2018 to May 2020,12 cases of congenital macrodactyly were divided into four groups of isolated macrodactyly of fingers,isolated macrodactyly of toes,macrodactyly and syndactyly of fingers,macrodactyly and syndactyly of toes.The pathological tissues and peripheral blood of all the patients were detected by whole exome sequencing.The four groups of detected mutated genes were intersected,and then Phenolyzer analysis was performed on the results of intersection to screen the pathogenic genes of macrodactyly.The exon sequencing results were further verified by Sanger sequencing to identify the mutated genes and loci of congenital macrodactyly.Monoclonal antibody immunohistochemical analysis was performed on the protein signaling pathway encoded by the mutated gene.The adipocytes of polydactyly children were used as control group for comparison,and the expression of target protein was observed.Results According to the results of whole exon sequencing synthesis biosynthesis analysis and Sanger verification,PIK3CA gene mutation was screened out to be the pathogenic gene of macrodactyly.The mutation loci were c.G1624A(p.E542K)and c.G1633A(p.E545K)of exon 10,c.A3140G(p.H1047R)and c.G3145C(p.G1049R)of exon 21,among which c.G3145C(p.G1049R)was first discovered.Immunohistochemical analysis of AKT monoclonal antibody showed that AKT protein expression was enhanced in macrodactyly.Conclusion The overexpression of PI3K-Akt signaling pathway caused by PIK3CA gene mutation is the cause of congenital macrodactyly.

关 键 词：手畸形 足畸形 巨指(趾) PIK3CA基因 

分 类 号：R658.2[医药卫生—外科学]