title: Investigation of Somatic NKX2-5 Mutations in Chinese Children with Congenital Heart Disease.
abstract: The purposes of this study are to investigate somatic NKX2-5 mutations in Chinese children with congenital heart disease (CHD) and assess the reliability of somatic mutation detection in formalin-fixed, paraffin-embedded (FFPE) tissues. The study cohort included frozen and FFPE cardiac tissues as well as blood samples from 85 Chinese children with CHD who had the cardiac operations. The right atrial appendage far from the diseased heart was used as normal control. Genomic DNA was isolated from cardiac tissues and blood samples using TIANamp Blood DNA kit. Two exons and exon-intron boundaries of NKX2-5 were amplified by polymerase chain reaction (PCR) and sequenced by dideoxynucleotide chain termination approach. The acquired sequences were aligned with GenBank sequences to identify the sequence variations. No somatic mutation in the NKX2-5 gene was observed in both frozen and FFPE cardiac tissues in 85 Chinese children with CHD. Nonetheless, a common single nucleotide polymorphism (SNP), c.63 A > G (E21E), was identified in all the three kinds of DNA samples with the same allele frequency 82.3%. Moreover, another common SNP c.606 G > C (L202L) was found in 2.3% of our patients. There were no significant differences in the allele frequencies of two SNPs between the cardiac diseased tissues and right atrial appendage (P > 0.05). PCR artefact as mutations was not found in the FFPE tissues stored for one year. Our findings demonstrate that somatic NKX2-5 mutations do not represent an important aetiologic pathway in Chinese children with congenital heart disease.
authors.affiliation: 1. Institute of Pediatric Translational Medicine, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China. ; 2. Department of Pediatric Cardiology, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Jiayi
authors.initials: J
authors.lastname: Zheng
authors.affiliation: 2. Department of Pediatric Cardiology, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Li
authors.affiliation: 3. Department of Cardiothoracic Surgery, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Jinfen
authors.initials: J
authors.lastname: Liu
authors.affiliation: 3. Department of Cardiothoracic Surgery, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Xu
authors.affiliation: 3. Department of Cardiothoracic Surgery, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Haibo
authors.initials: H
authors.lastname: Zhang
authors.affiliation: 1. Institute of Pediatric Translational Medicine, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China. ; 4. Clinical Laboratory, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Qihua
authors.initials: Q
authors.lastname: Fu
authors.affiliation: 5. Molecular Diagnostics Laboratory, Shanghai Children's Medical Center Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: 6. Pediatric Cardiology Department, Xinhua Hospital Affiliated to School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Sun
mesh_terms: D044466:Asian People; D006348:Cardiac Surgical Procedures; D002675:Child, Preschool; D002681:China; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D006330:Heart Defects, Congenital / Q000097:blood / Q000235:genetics* / Q000473:pathology; D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins / Q000097:blood / Q000235:genetics*; D006801:Humans; D007223:Infant; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D014157:Transcription Factors / Q000097:blood / Q000235:genetics*
chemical_list: D000072576:Homeobox Protein Nkx-2.5; D018398:Homeodomain Proteins; C095552:NKX2-5 protein, human; D014157:Transcription Factors
keywords: Congenital Heart Disease (CHD); NKX2-5; formalin-fixed; paraffin-embedded (FFPE) tissues; single nucleotide polymorphisms (SNP); somatic mutation