目的：了解 GATA-4，-5，-6基因突变与先天性心脏病（CHD）的关系，为 CHD 患儿的早期预防及遗传咨询提供支持。方法收集室间隔缺损患儿66例、房间隔缺损患儿84例及单纯性心脏圆锥动脉干畸形患儿48例的临床资料和外周静脉血样本，共198例 CHD 患儿；并以300例健康者为健康对照。应用聚合酶链反应（PCR）扩增 GATA-4，-5，-6基因的全部外显子和两侧部分内含子，PCR 产物纯化后以自动测序仪正反向测序。应用 BLAST 程序将所测 GATA-4，-5，-6基因序列与 GenBank 中已知的序列进行比对，检测基因突变。结果在1例室间隔缺损患儿和1例单纯性心脏圆锥动脉干畸形即永存动脉干畸形患儿中发现转录因子GATA-4相同的杂合子突变，第4外显子区 c．799G ＞A，突变使转录因子蛋白第267位的缬氨酸被蛋氨酸取代即p．V267M，多物种氨基酸序列比对结果显示该突变氨基酸在进化上高度保守。转录因子 GATA-5，-6在本组CHD 患儿中筛查未发现突变。结论CHD 发生机制复杂，转录因子 GATA-4突变与人类 CHD 的发生相关，转录因子 GATA-4可能是人类 CHD 的易感基因。%Objective To understand the relationship between GATA -4,-5,-6 gene mutations and con-genital heart disease(CHD),and to provide grounds for early prevention and genetic counseling of children with CHD. Methods GATA -4,-5,-6 coding regions exons and the flanking intron sequences in 1 98 CHD patients were screened,including 66 cases of the ventricular septal defects,84 cases of the atrial septal defects,and 48 cases of the nonsyndromic conotruncal heart defects patients.A total of 300 healthy subjects were selected as controls.The acquired sequences were aligned with which those publicized in GenBank by the aid of program BLAST.All exons and bilateral partial intron -exon boundaries of GATA -4,-5,-6 genes were amplified by the polymerase chain reaction (PCR). The PCR products were purified and directly sequenced by automatic DNA sequence equipment.The acquired GATA -4,-5,-6 gene sequences were compared with GenBank standard gene sequences with the aid of program BLAST. Results A heterozygous missense mutation in the GATA -4 gene was identified in a ventricular septal defect patient and a persistent truncus arteriosus patient.The mutation was located in c.799G >A(p.V267M)in exon 4 of GATA -4. Multiple aligenment of GATA -4 proteins across species demonstrated that altered amino acid was highly conserved. Transcription factor GATA -5,-6 screening showed no mutations in children with CHD in this study.Conclusions Transcription factor GATA -4 gene mutation may be associated with the occurrence of CHD.Transcription factor GATA -4 gene may be susceptible gene in human CHD.
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