12家系视网膜母细胞瘤基因诊断的回顾性分析  

Retrospective Study on Gene Diagnosis of Retinoblastoma in 12 Families

作　　者：白周现 张翠云 邵敬芝[3] 刘莉娜 孔祥东[1] Bai Zhouxian;Zhang Cuiyun;Shao Jingzhi;Liu Lina;Kong Xiangdong(The Genetics and Prenatal Diagnosis Center,the Department of Obstetrics and Gynecology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;The Department of Molecular Pathology,Affiliated Cancer Hospital of Zhengzhou University,Henan Cancer Hospital,Zhengzhou 450000,China;The Department of Ophthalmology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学第一附属医院妇产科遗传与产前诊断中心,450000 [2]郑州大学附属肿瘤医院河南省肿瘤医院分子病理科,450000 [3]郑州大学第一附属医院眼科,450000

出　　处：《中华眼视光学与视觉科学杂志》2021年第11期820-826,共7页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：国家重点研发计划子课题(2018YFC1002206-2);郑州大学第一附属医院院内青年创新基金(YNQN2017008)。

摘　　要：目的: 检测并分析视网膜母细胞瘤患者的遗传学病因, 为患者及其家系基因诊断、遗传咨询、产前诊断提供理论依据。方法: 实验研究。收集12个家系视网膜母细胞瘤患者的临床检查资料, 采集患者及家系受检者外周血提取基因组DNA, 已故患者由家属提供石蜡切片组织提取基因组DNA, 抽取相关孕妇羊水细胞提取基因组DNA。应用二代测序筛查、Sanger测序法验证RB1基因候选变异。通过相关数据库和PubMed文献检索相关变异的致病性, 依据遗传变异分类标准指南判断并分级候选变异的致病性。依据家系先证者致病突变, 相关孕妇行RB1基因产前诊断。结果: 纳入的12个视网膜母细胞瘤家系, 其中8个家系检测到致病基因变异, 检测阳性率67%。检测到的8个不同RB1基因突变中, 6个为已报道致病性变异;RB1基因c.2404dupG(p.N803Efs*12)和c.1380;381insCTTA(p.K462Tfs*2)为本研究发现的新突变。变异致病性综合分析本研究检测到的8个不同RB1基因变异均为致病性。结论: 本研究纳入的12个视网膜母细胞瘤家系中有8个家系患者检测到致病突变而获得基因诊断;本研究同时发现了导致视网膜母细胞瘤的新突变。相关家系依据基因诊断结果进行了有效的遗传咨询和产前诊断。Objective:To detect and analyze the genetic etiology of retinoblastoma patients,and to provide a theoretical basis for gene diagnosis,genetic counseling and prenatal diagnosis of patients and their families.Methods:This was an experimental study.Clinical material from 12 families were collected and genomic DNA was extracted from the peripheral blood of patients and related subjects.An FFPE sample was collected from adeceased patient and genomic DNA was extracted.Genomic DNA was extracted from the amniotic fluid cells of the pregnant women.The mutation was screened by using targeted sequencing,and PCR-Sanger sequencing was used to verify the selected mutations.PubMed and related databases were searched.Pathogenic analysis of the mutation and the mutation effect was interpreted by protein prediction software.The pathogenicity of candidate mutations was determined and graded according to a joint consensus recommended by the American College of Medical Genetics and Genomics and the Association for Molecular Pathology.According to the pathogenic mutation of the proband in the family,the related pregnant women underwent RB1 gene prenatal diagnosis.Results:Different pathogenic gene variations,6 of which were known pathogenic variations,were detected in 8 of the 12 selected families,and the positive rate was 67%.And the RB1 gene c.2404dupG(p.N803Efs*12)and c.1380_1381insCTTA(p.K462Tfs*2)detected in this study were novel genes.All of them were pathogenic based on comprehensive analysis of the eight different RB1 gene mutations detected in this study.Conclusions:Eight of the twelve families with retinoblastoma were diagnosed with RB1 mutation.The study found two new mutations in retinoblastoma.Based on the results of the genetic diagnosis,the related families received effective genetic counseling and prenatal diagnosis.

关 键 词：视网膜母细胞瘤 RB1基因 靶向测序 产前诊断 

分 类 号：R739.7[医药卫生—肿瘤]