转录因子NKX2.5和GATA4基因多态性与6个先天性心脏病室间隔缺损家系的相关性研究

发布时间：2019-06-12 15:11

【摘要】： [目的] 先天性心脏病(Congenital heart disease, CHD)是人类所有出生缺陷中最为常见的疾病之一,是导致婴幼儿残疾和死亡的重要原因。室间隔缺损(ventricular septal defect, VSD)占新生儿CHD发病总数的20%,占儿童CHD发病总数的30%,是所有CHD中发病率最高的一种。而调控心脏发育的基因若发生突变会导致CHD的发生,因此寻找VSD发生的相关基因成为心血管分子遗传学领域研究焦点之一。目前在众多的基因当中,国内外研究最多的是参与正常心脏发育过程中的转录因子NKX2.5和GATA4,它们在心脏发育过程中进行着不同的时间和空间表达,从而调节心脏的正常发育,任何影响它们正常表达的因素均有可能导致心脏畸形。本研究从VSD家系的角度探讨NKX2.5和GATA4基因多态性和CHD的关系,期望为进一步阐明CHD家系发病的分子机理提供新的实验依据。 [方法]本研究采用云南省心血管病医院心胸外科采集的6个室间隔缺损家系,包括10名VSD患者和20名健康成员的血样标本,并根据其临床资料建立家系系谱图并提取血样标本DNA,利用PCR扩增技术和DNA直接测序的方法对NKX2.5和GATA4基因的部分外显子及3'UTR区域的多态性进行检测,并对各个位点的多态与VSD的相关性进行分析。 [结果] 1.初步建立了中国云南地区部分室间隔缺损家系的临床信息及DNA库； 2.在VSD患者及正常人群中均检测到NKX2.5基因外显子1上的rs2277923突变位点的多态; 3.仅在家系Ⅲ的父子两名患者中发现NKX2.5基因3’端非翻译区的rs703752位点的一个CA的突变； 4.GATA4基因外显子6检测的单核苷酸多态性位点均未发现突变点。 [结论] 1.已收集到的6个VSD家系的临床资料丰富了VSD家系的遗传资源信息； 2.NKX2.5基因外显子1上的rs2277923位点检测到多态,但是我们认为该同义突变位点与CHD发病的相关性仍然需要进一步的深入研究； 3.首次在国内对NKX2.5基因rs703752位点的多态进行报道,且研究结果提示该位点的突变与CHD的发病有关； 4.本课题研究的结果,对于该领域的进一步研究具有重要的参考价值,为以后的基因筛查和基因诊断具有指导意义。
[Abstract]:[objective] congenital heart disease (Congenital heart disease, CHD) is one of the most common diseases in all human birth defects, and it is an important cause of disability and death in infants and young children. Ventricular septum defect (ventricular septal defect, VSD) accounted for 20% of the total incidence of neonatal CHD and 30% of the total incidence of CHD in children. It was the highest incidence of CHD among all CHD. However, the mutation of genes regulating cardiac development will lead to the occurrence of CHD, so finding the genes related to the occurrence of VSD has become one of the research focuses in the field of cardiovascular molecular genetics. At present, among many genes, the transcription factors NKX2.5 and GATA4, which are involved in normal cardiac development, are expressed in different time and space during cardiac development, thus regulating the normal development of heart. Any factors affecting their normal expression may lead to cardiac malformation. In this study, the relationship between NKX2.5 and GATA4 gene polymorphism and CHD was discussed from the point of view of VSD pedigree, and it was expected to provide a new experimental basis for further elucidating the molecular mechanism of CHD family. [methods] the blood samples of 6 families with ventricular septal defect (VSD), including 10 VSD patients and 20 healthy members, were collected from cardiothoracic surgery department of Yunnan Cardiovascular Hospital. The pedigree maps were established according to their clinical data, and the DNA, samples were extracted to detect the polymorphism of some exons and 3'UTR regions of NKX2.5 and GATA4 genes by PCR amplification and DNA direct sequencing. The correlation between polymorphism and VSD at each site was analyzed. [result] 1. The clinical information and DNA database of some families with ventricular septal defect (VSD) in Yunnan Province of China were established. The polymorphism of rs2277923 mutation site in Exon 1 of NKX2.5 gene was detected in VSD patients and normal population. Only one CA mutation at the rs703752 site in the untranslated region of the 3 'end of the NKX2.5 gene was found in the father and son of the family III, and no mutation point was found in the single nucleotides polymorphism site detected in the exon6 of the 4.GATA4 gene. [conclusion] 1. The clinical data of six VSD families have enriched the genetic resource information of VSD families, and the polymorphism of rs2277923 site in 2.NKX2.5 gene exon1 has been detected, but we think that the correlation between the synonymous mutation site and the pathogenesis of CHD still needs further in-depth study. 3. The polymorphism of rs703752 site of NKX2.5 gene was reported for the first time in China, and the results suggested that the mutation of this site was related to the pathogenesis of CHD. The results of this study have important reference value for further research in this field, and have guiding significance for gene screening and gene diagnosis in the future.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R346

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