先天性心脏病相关PITX2c基因新突变研究  

Study of a novel PITX2 cmutation associated with congenital heart disease

作　　者：潘云[1] 周宁[1] 赵红[1] 郑桂芬[1] 刘兴元[1] 

机构地区：[1]同济大学医学院附属同济医院儿科,上海200065

出　　处：《国际心血管病杂志》2015年第3期194-197,共4页International Journal of Cardiovascular Disease

基　　金：上海市自然科学基金(10ZR1433100)

摘　　要：目的:研究先天性心脏病(congenital heart disease,CHD)相关PITX2c基因的新突变。方法:收集150例CHD患者和200名正常对照者的外周静脉血标本,使用DNA纯化试剂盒分离基因组DNA。使用DNA聚合酶扩增PITX2c基因的编码区和剪接位点,应用DNA测序试剂盒在DNA分析仪上对扩增片段进行测序。将所测序列与GenBank数据库中的PITX2c基因序列进行比对以发现PITX2c基因突变。使用在线程序MUSCLE分析突变氨基酸的保守性,分别应用MutationTaster和PolyPhen-2分析突变氨基酸的致病性。结果:在1例散发性CHD患者发现了1个新的PITX2c基因杂合错义突变,即p.S101G突变,突变率约为0.67%。该错义突变不存在于200名正常对照者。跨物种PITX2c蛋白之氨基酸序列对比显示第101位的丝氨酸在进化上完全保守,致病性预测显示所发现的PITX2c基因变异是致病性突变。结论:本研究揭示了CHD相关PITX2c基因新突变,对于制定新的CHD防治策略具有潜在的意义。Objective：To study a novel PITX2 cmutation associated with congenital heart disease（CHD）. Methods：Clinical data and peripheral venous blood specimens from 150 patients with CHD and 200 healthy individuals used as controls were collected.The genomic DNA was isolated with DNA extraction kit.The coding regions and splicing sites of the PITX2 cgene were amplified by using DNA polymerase.The amplified products were sequenced with DNA sequencing kit under DNA Analyzer.The obtained sequences were aligned with those of PITX2 cfrom GenBank to identify a novel PITX2 c variation.The online program MUSCL was used to analyze whether the altered amino acid was conserved evolutionarily.MutationTaster and PolyPhen-2 were used to predict the disease-causing likeliness,respectively. Results：A novel heterozygous PITX2 cmutation,p.S101 G,was identified in a patient with sporadic CHD,with a mutational prevalence of about 0.67%.The missense mutation was absent in the 200 control subjects.Multiple alignments of PITX2 cproteins across species exhibited that the altered amino acid was completely conserved evolutionarily.The identified PITX2 cvariation was predicted to be apathogenic mutation. Conclusion：This study reveals a novel PITX2 cmutation associated with CHD,suggesting potential implications in the development of new prophylactic and therapeutic strategies for CHD.

关 键 词：先天性心脏病 室间隔缺损 遗传学 转录因子 PITX2c 

分 类 号：R725.4[医药卫生—儿科]