Screening Of TFAP2B Gene Mutation In Children With Patent Ductus Arteriosus

ObjectivesPatent ductus arteriosus (PDA) is a common congenital heart disease in children. Genetic factor is one of the important factors in the occurrence of PDA, so searching for the related genes of PDA has been one of the focuses in the field of cardiovascular diseases. Cardiac-specific transcription factor TFAP2B plays an important role in neural crest cell differentiation and the formation of the ductus arteriosus. This study was sought to investigate whether there was any mutation in TFAP2B gene in PDA children.MethodsA total of74children with simple PDA was used in the experiment, which were confirmed by color Doppler echocardiography for simple PDA, and collected a total of100healthy children as controls. Genomic DNA was extracted from whole blood samples. Use primer premier5.0software to design seven primers of TFAP2B gene and then amplified all exons and flanking intronic regions by polymerase chain reaction. PCR fragments were purified after agarose gel electrophoresis, then sequenced using the Big-Dye Termination Cycle Sequencing kit on an ABI PRIME3730DNA sequencer. The required sequences were compared to NCBI and Genbank sequence databases, then analysis whether there was any gene mutation or polymorphism.Results 1. In sequencing of all exons of TFAP2B gene in children with PDA, non-synonymous mutations were not found.2. In sequencing of TFAP2B gene from74cases of children with PDA, one novel single nucleotide polymorphism (c.1-34G→A) was identified on the34point of TFAP2B gene coding sequence in26cases. Equally, this SNP was also identified in32cases of100controls, which was not included in NCBI and Genbank sequence databases. The frequency distribution on this point of the three genotypes of the GG, AG, AA in the overall population is according with Hardy-Weinberg genetic equilibrium (x=0.129, P=0.938). However, no significant difference was observed in frequency distributions of the alleles and genotypes between children with PDA and control subjects (P>0.05).Conclusions1. None mutation was found in the sequencing of TFAP2B gene in children with PDA.2. In the study, one novel single nucleotide polymorphism (c.1-34G→A) was identified on the34point of TFAP2B gene coding sequence in26cases of74children with PDA and32cases in100healthy children, it suggests that the novel single nucleotide polymorphism of the TFAP2B gene may be not related to PDA.