Nkx2-5在单纯性先天性心脏病中的突变及表达研究

Study on the mutation and expression of Nkx2-5 gene in human simple congenital heart disease

目的分析Nkx2-5基因在单纯性先天性心脏病(congenital heart disease,CHD)心肌组织中的突变及表达情况,探讨Nkx2-5基因突变、表达与单纯性CHD发生机制的关系。方法采用PCR-SSCP(单链构象多态性)方法对30例因CHD引产胎儿心脏组织进行Nkx2-5基因编码序列突变筛查;以β-actin为内对照,用RT-PCR方法检测Nkx2-5基因在单纯性CHD引产胎儿心肌组织中mRNA的表达情况。结果 30例单纯性CHD引产胎儿Nkx2-5基因2个外显子PCR产物经SSCP检测未发现突变;与正常对照心肌组织相比,单纯性CHD胎儿该基因mRNA表达呈下降趋势(P<0.05)。结论单纯性CHD中Nkx2-5基因编码区的体细胞突变可能不是单纯性CHD的致病机制;Nkx2-5基因转录水平异常可能是该基因参与CHD形成的一种潜在机制。

Objective：We investigated the mutation and expression of Nkx2-5 gene in human simple Congenital Heart Disease（CHD） so as to clear relationship of the mutation and expression and simple CHD.Methods：We examine the mutations of coding sequence of Nkx2-5 gene in 30 myocardium samples from fetuses with simple CHD by PCR single strand conformation polymorphism（PCR-SSCP）;Using β-actin as internal control,we detected the differential expression between 30 myocardium samples from simple congenital heart disease fetuses and 30 normal controls by reverse transcription polymerase chain reaction（RT-PCR）.Results：No mutation was detected in 2 exons of Nkx2-5 gene in all samples by PCR-SSCP;The mRNA expression levels of Nkx2-5 show descent tendency in samples of simple CHD compared with normal controls.Conclusion：Mutations in coding region of Nkx2-5 gene may not have the somatic cell character,the abnormality in transcription level of Nkx2-5 gene may be a kind of mechanism causing human simple CHD.