【摘要】 目的研究心肌特异性转录因子NKX2-5基因启动子区在先天性心脏病圆锥动脉干畸形(CTD)病例中的甲基化状态,从表观遗传学角度探讨先心病的发病机制,为先心病的早期临床诊治提供理论依据。方法首先利用Tru1l限制性内切酶将基因组片段化并接上接头,然后利用甲基化结合蛋白(MBD)与甲基化DNA特异性结合的特点对甲基化DNA片段进行富集;针对NKX2-5基因启动子区的CpG岛设计引物,通过实时定量PCR检测CTD与正常胎心组织中NKX2-5基因甲基化状态。结果通过与正常心脏组织相比,NKX2-5基因在6例病变组织中均发生了不同程度的甲基化,其中4例呈现高甲基化,另外2例甲基化程度无明显变化。结论 NKX2-5基因甲基化可能与先心病CTD发病相关,可能成为先心病早期诊断的分子标志物之一。更多还原

【Abstract】 Objective To study the promoter methylation status of heart tissue-specific transcription factor NKX2-5 gene between congenital heart conotruncal defects (CTD) and normal fetal cardiac tissue samples,and to investigate the pathogenesis of congenital heart disease from the epigenetic point of view. Methods First,the genome DNA fragments were generated by the Tru1l restriction enzyme and then connected to the adaptor. The methylated DNA fragments were enriched by the methylated binding protein (MBD),which can specifically bind and interact with methyl-CpG island. Finally,the CpG island methylation status in the promoter region of NKX2-5 gene was detected by Real Time PCR.Results In 6 CTD cases we detected,4 showed hypermethylation status in NKX2-5 gene promoter,compared with normal fetal heart tissue.Conclusion NKX2-5 gene hypermethylation might be one of molecular marker of congenital heart disease.更多还原