中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2008年
6期
637-641
,共5页
齐秋锋%易龙%杨驰%陈慧梅%沈力%莫绪明%胡娅莉%王亚平
齊鞦鋒%易龍%楊馳%陳慧梅%瀋力%莫緒明%鬍婭莉%王亞平
제추봉%역룡%양치%진혜매%침력%막서명%호아리%왕아평
CHD7基因%先天性心脏病%变性高效液相色谱%突变
CHD7基因%先天性心髒病%變性高效液相色譜%突變
CHD7기인%선천성심장병%변성고효액상색보%돌변
CHD7 gene%congenital heart disease%denaturing high performance liquid chromatography%mutationK200605,supported by the Foundation for Key Project of Jiangsu Health Department
目的 筛查先天性心脏病(congenital heart fflsease,CHD)患者CHD7(chromodomsin helicase DNA-binding protein gene)基因的胚系突变,探讨其在先心病发生中的作用.方法 采集67例临床确诊的CHD患儿、100名正常对照的外周静脉血,提取白细胞基因组DNA,PCR扩增CHD7基因,变性高效液相色谱分析技术对PCR产物进行突变筛选,出现异常峰型的扩增片段进行DNA测序,明确突变位点和类型,并进一步结合病例对照和生物信息学分析探讨变异的功能意义.结果 67例先心病患者中共检出7种单碱基的替换,位于CHD7的不同内含子.其中,IVS11+127A＞G和WS12+21T＞G的等位基因频率均为0.0075,为罕见变异;而IVS2+34G＞A、IVS4+39C＞A、IVS12.5T＞C和IVS16+51C＞A等位基因频率为0.2635、0.2156、0.1505、0.3636,属于单核苷酸多态性;IVS12-5T＞C在CHD组的检出频率显著低于正常对照组(5.42%vs 9.57%,P＜0.05);而IVS14-35C＞G则仅见于CHD患儿.生物信息学分析显示,IVS12-5T＞C替换具有增强外显子剪切的效应.结论 CHD7基因单核苷酸多态性变异WS12-5T＞C对CHO的发生可能具有保护作用,而CHD7基因的突变并不是构成散发性先天性心脏病的主要原因.
目的 篩查先天性心髒病(congenital heart fflsease,CHD)患者CHD7(chromodomsin helicase DNA-binding protein gene)基因的胚繫突變,探討其在先心病髮生中的作用.方法 採集67例臨床確診的CHD患兒、100名正常對照的外週靜脈血,提取白細胞基因組DNA,PCR擴增CHD7基因,變性高效液相色譜分析技術對PCR產物進行突變篩選,齣現異常峰型的擴增片段進行DNA測序,明確突變位點和類型,併進一步結閤病例對照和生物信息學分析探討變異的功能意義.結果 67例先心病患者中共檢齣7種單堿基的替換,位于CHD7的不同內含子.其中,IVS11+127A＞G和WS12+21T＞G的等位基因頻率均為0.0075,為罕見變異;而IVS2+34G＞A、IVS4+39C＞A、IVS12.5T＞C和IVS16+51C＞A等位基因頻率為0.2635、0.2156、0.1505、0.3636,屬于單覈苷痠多態性;IVS12-5T＞C在CHD組的檢齣頻率顯著低于正常對照組(5.42%vs 9.57%,P＜0.05);而IVS14-35C＞G則僅見于CHD患兒.生物信息學分析顯示,IVS12-5T＞C替換具有增彊外顯子剪切的效應.結論 CHD7基因單覈苷痠多態性變異WS12-5T＞C對CHO的髮生可能具有保護作用,而CHD7基因的突變併不是構成散髮性先天性心髒病的主要原因.
목적 사사선천성심장병(congenital heart fflsease,CHD)환자CHD7(chromodomsin helicase DNA-binding protein gene)기인적배계돌변,탐토기재선심병발생중적작용.방법 채집67례림상학진적CHD환인、100명정상대조적외주정맥혈,제취백세포기인조DNA,PCR확증CHD7기인,변성고효액상색보분석기술대PCR산물진행돌변사선,출현이상봉형적확증편단진행DNA측서,명학돌변위점화류형,병진일보결합병례대조화생물신식학분석탐토변이적공능의의.결과 67례선심병환자중공검출7충단감기적체환,위우CHD7적불동내함자.기중,IVS11+127A＞G화WS12+21T＞G적등위기인빈솔균위0.0075,위한견변이;이IVS2+34G＞A、IVS4+39C＞A、IVS12.5T＞C화IVS16+51C＞A등위기인빈솔위0.2635、0.2156、0.1505、0.3636,속우단핵감산다태성;IVS12-5T＞C재CHD조적검출빈솔현저저우정상대조조(5.42%vs 9.57%,P＜0.05);이IVS14-35C＞G칙부견우CHD환인.생물신식학분석현시,IVS12-5T＞C체환구유증강외현자전절적효응.결론 CHD7기인단핵감산다태성변이WS12-5T＞C대CHO적발생가능구유보호작용,이CHD7기인적돌변병불시구성산발성선천성심장병적주요원인.
Objective To investigate the germline mutations of the CHD7 gene and their roles in patients with congenital heart disease (CHD).Methods Genomic DNAs extracted from peripheral blood were subjected to screen mutations in CHD7 germ by denaturing high performance liquid chromatography (DHPLC) followed by DNA sequencing of aberrant peaks in 67 CHD patients and 100 healthy control.Case-control study and bioinformatie analysis were utilized to explore the potential functional roles of the variations detected.Resells Seven kinds of single nucleotide substitution were detected in the CHD patients in different introus of the CHD7 gene.Among them,IVS11+127A＞G and IVS12 +21T＞ G were rare variations and the allele frequencies of both were 0.0075;Mille IVS2 + 34G＞A,IVS4 + 39G＞A,IVS12-5T＞ C and IVS16 + 51C＞ A were the single nucleotide polymorphisms and the allele frequency was 0.2635,0.2156,0.1505 and 0.3636 respectively.The frequency of IVS12-5T＞ C in the CHD group was significantly lower than that in the control group (5.42% versus 9.57%,P ＜0.05).The variant of IVS14-35C＞G was only detected in patients with CHD.Bioinformatie analysis showed that IVS12-5T＞C might increase exon splicing ability comparing with the wild-type sequence.Conclusion The CHD7 germ mutation may not be the main reason for sporadic congenital heart disease,whereas the single nucleotide polymorphism of IVS12-5T＞C might play a protective role in the onset of this dis-ease.
