动脉导管未闭患儿TFAP-2B基因突变的研究  

Mutation of TFAP-2B gene in patients with patent ductus arteriosus

作　　者：陈轶维[1] 赵武[3] 李奋[1] 吉炜[1] 傅启华[2] 张志芳[1] 王剑[1] 

机构地区：[1]上海交通大学医学院附属上海儿童医学中心心血管内科,200127 [2]上海交通大学医学院附属上海儿童医学中心检验科,200127 [3]蚌埠医学院附属第一医院儿科

出　　处：《中华儿科杂志》2010年第12期900-904,共5页Chinese Journal of Pediatrics

基　　金：基金项目：国家十一五科技支撑计划（2007BAl05803）;上海高校创新团队发展计划资助（沪教委科2010.29号文）

摘　　要：目的 发现我国动脉导管未闭（patent ductus arteriosus,PDA）患儿分子遗传方面缺陷,为PDA早期预防及遗传咨询提供支持.方法 收集100例单纯性PDA患儿的临床资料和外周静脉血样本,以100名健康儿童为对照.应用聚合酶链式反应（polymerase chain reaction,PCR）扩增TFAP-2B基因的全部外显子和外显子两侧部分内含子,并对扩增片段进行双向测序.应用BLAST程序将所测TFAP-2B基因序列与GeneBank中的已知序列进行对比以检测基因突变.采用逆转录聚合酶链式反应（reverse transcription polymerase chain reaction,RT-PCR）对1例家族史阳性患儿及其家属共16人TFAP-2B部分cDNA片段进行扩增,扩增产物直接进行双向序列测定.结果 基因分析显示,在1例家族史阳性患儿及其患病亲属中,TFAP-2B第3内含子剪接位点＋5位发生突变[intron3（＋5）G＞A],患儿TFAP-2B基因部分cDNA巢式PCR扩增结果 提示3号外显子完全缺失.此外,还发现了一个新的单核苷酸多态性,即转录起始点上游第34位的鸟嘌呤变为腺嘌呤,这个多态在PDA患者和健康对照组的频率分布差异有统计学意义（Z=-2.513,P=0.012）.结论 TFAP-2B基因突变能够导致家族型PDA.Objective To identify novel genetic mutations in Chinese patients with congenital patent ductus arteriosus （PDA）. Method Clinical data and peripheral blood specimens from a kindred spanning 3 generations in which 5 of 16 individuals had PDA and a cohort of 95 unrelated subjects with PDA were collected, and 100 unrelated healthy individuals were included as controls. The coding exons and flanking introns of TFAP-2B gene were amplified by polymerase chain reaction （ PCR ） with specific primers. We aligned the acquired sequences with which publicized in GenBank by the aid of program BLAST. Reverse transcription-polymerase chain reaction （RT-PCR） was used to amplify the parts of TFAP-2B and sequencing was performed on PCR products forward and reversely directly. Result Sequencing of TFAP-2B identified that there was a splice-junction in intron 3 [ intron3 （ ＋ 5 ） G 〉 A ] and a 60 bp deletion was found in exon 3 by nested PCR. Additional]y, a novel single nucleotide polymorphism （SNP） where a transition of guanine （G） to adenine （A） was identified at 34 bp front of transcription initiation site in TFAP-2B gene. There were significant differences in the prevalence of alleles G and A between controls and PDA patients （ Z =-2.513,P= 0.012）. Conclusion We identified a novel splice-junction in TFAP-2B gene which might lead to hereditary PDA in a Chinese family. However, the mechanism by which this mutation results in PDA is still to be ascertained.

关 键 词：动脉导管未闭 转录因子AP-2 突变 

分 类 号：R725.4[医药卫生—儿科]