单倍型dce中RHCE*ce等位基因变异致e抗原减弱1例的分子生物学研究  

Molecular study of a case with variant of RHCE*ce allele in haplotype dce resulting in weakened e antigen

作　　者：孔永奎[1] 杨贺才[2] 邵明[1] 陈李影慧[1] 张婉金 张小燕 王静[1] 吕先萍[1] 杨乾坤[1] Kong Yongkui;Yang Hecai;Shao Ming;Chen-Li Yinghui;Zhang Wanjin;Zhang Xiaoyan;Wang Jing;Lyu Xianping;Yang Qiankun(Department of Blood Transfusion,the First Affiliated Hospital of Zhengzhou University,Zhengzhou,Henan 450052,China;Department of Blood Group Research,Henan Red Cross Blood Center,Zhengzhou,Henan 450014,China)

机构地区：[1]郑州大学第一附属医院输血科,郑州450052 [2]河南省红十字血液中心血型研究室,郑州450014

出　　处：《中华医学遗传学杂志》2024年第9期1039-1044,共6页Chinese Journal of Medical Genetics

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20220260)。

摘　　要：目的探讨1例RhD(-)女性先证者的RH基因型及其分子生物学机制。方法选取2019年8月就诊于郑州大学第一附属医院的1例26岁女性先证者作为研究对象。采集先证者及其父母的外周血样,对其进行卡式法Rh表型检测。用PCR-基于序列法(PCR-sequence-based typing,PCR-SBT)和基因测序法检测先证者及其父母的RHD合子型和RH基因型。用生物信息学软件进行Rh蛋白同源建模,并通过分子动力学模拟变异所造成的蛋白结构改变。本研究通过郑州大学第一附属医院医学伦理委员会的审查(伦理号:2023-KY-0870-003)。结果血清学检测显示先证者及其父亲Rh分型e抗原减弱。PCR-SBT及基因测序显示3人的基因型分别为dce/dCE、dce/DcE、dCE/DcE,先证者RHD和RHCE的基因型分别为RHD*01N.01/RHD*01N.16、RHCE*01.01/RHCE*04。蛋白模拟和分子动力学分析显示RHCE*ce(c.48G>C)所导致的ce_16C变异将造成膜内外环结构的改变,主要影响膜外第2、6环和膜内第3、4的摆动性。结论RHCE*ce等位基因的c.48G>C变异可能是导致先证者e抗原减弱的原因。ObjectiveTo explore the RH genotype for a female with RhD(-)blood type and its molecular basis.MethodsA 26-year-old female who had attended the outpatient clinic of the First Affiliated Hospital of Zhengzhou University in August 2019 was selected as the study subject.Peripheral blood samples were collected from the proband and her parents for Rh phenotyping with gel card method.PCR-sequence-based typing(PCR-SBT)and DNA sequencing were used to determine the RHD zygosity and RH genotype of the proband and her parents.Homology modeling of Rh proteins was performed with bioinformatic software,and protein structural alterations caused by the variant was simulated by molecular dynamics.This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University(Ethics No.2023-KY-0870-003).ResultsSerological tests showed that the proband and her father both had weakened e antigen of the Rh phenotype.PCR-SBT and DNA sequencing showed that the genotypes of the proband and her parents were dce/dCE,dce/DcE and dCE/DcE,respectively.And the genotypes of the RHD and RHCE of the proband were RHD*01N.01/RHD*01N.16,RHCE*01.01/RHCE*04,respectively.Protein simulation and molecular dynamics analysis revealed that the ce_16C variant resulted from RHCE*ce(c.48G>C)may alter the structure of intracellular and extracellular loops,mainly affecting the mobility of extracellular loops 2,6 and intracellular loops 3,4.ConclusionVariant of the RHCE*ce allele c.48G>C probably underlay the weakened e antigen in this proband.

关 键 词：RH血型 RH基因型 单倍型 同源建模 分子动力学 

分 类 号：R596[医药卫生—内科学]