上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

心脏圆锥动脉干畸形患儿TBX1C基因3′UTR区域突变分析

方绍海1，徐月娟1，曹瑞雪1，陈 笋1，徐 让2   

  1. 上海交通大学 医学院附属新华医院 1.小儿心血管科， 2.科研中心， 上海 200092
  • 出版日期:2014-06-28 发布日期:2014-06-30
  • 作者简介:方绍海（1988—）， 男， 硕士生; 电子信箱: fangshaohai1988@163.com。
  • 基金资助:

    国家自然科学基金（81070135）; 国家重点基础研究发展计划（“973”计划）（2010CB529501）;上海高校一流学科B类资助

Analysis of mutation of 3′UTR region of TBX1C gene for children with conotruncal heart defects

FANG Shao-hai1, XU Yue-juan1, CAO Rui-xue1, CHEN Sun1, XU Rang2   

  1. 1.Department of Pediatric Cardiology, 2.Scientific Research Center, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2014-06-28 Published:2014-06-30
  • Supported by:

    National Natural Science Foundation of China, 30800410；National Basic Research Program of China, “973” Program，2010CB529501;Shanghai College Leading Discipline Foundation B

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摘要：

目的 筛查心脏圆锥动脉干畸形(CTHD)患者中TBX1C基因3′UTR区域存在的基因突变，探讨该区域突变影响TBX1C基因表达的可能机制。方法 选取无22q11-2区微缺失的52例CTHD患儿（CTHD组）和89名健康儿童（对照组）作为研究对象，PCR扩增TBX1C基因完整3′UTR区序列，所有扩增片段均进行双向测序；将测序结果与GenBank中TBX1C 3′UTR序列（NM 080647.1）进行比对，筛查出可能存在的基因突变；PicTar和TargetScan在线预测软件预测可能与TBX1C基因3′UTR区结合的微RNA (miRNA)，分析TBX1C基因3′UTR区突变对miRNA调控TBX1C基因表达的影响。结果 在CTHD组中发现1例突变c.*164_*165insC，即在距离终止密码子164位与165位核苷酸之间插入1个胞嘧啶；该突变在对照组中不存在，其他研究中未见报道，为新发突变；预测结果显示10种miRNA能与TBX1C基因3′UTR区结合，该突变并不位于10 种miRNA与TBX1C3′UTR的结合区域。结论 TBX1C基因3′UTR区域存在新发突变c.*164_*165insC，该突变可能改变TBX1C3′UTR区域与miRNA结合的空间构象，进而影响miRNA对TBX1C基因表达的调控作用。

关键词: 圆锥动脉干畸形, TBX1基因, 基因突变, 3′UTR, 微RNA

Abstract:

Objective To screen mutations in 3′UTR region of TBX1C gene of patients with conotruncal heart defects (CTHD) and explore the possible mechanism that how mutations in this region affect the expression and function of TBX1C gene. Methods Fifty-two CTHD patients without 22q11.2 microdeletion (the CTHD group) and eighty nine healthy children (the control group) were selected. Sequences of 3′UTR region of TBX1C gene were amplified by the PCR and all PCR products were sequenced by both forward and reverse sequencing. Sequencing results were compared to TBX1C 3′UTR sequences in GenBank (NM 080647.1) and possible mutations were screened. Online software PicTar and TargetScan were used to predict microRNA (miRNA) which could bind to 3′UTR region of TBX1C gene. The effect of mutations in 3′UTR region of TBX1C gene on the expression of miRNA-regulated TBX1C gene was analyzed. Results A mutation of c.*164_*165insC was found in patients of the CTHD group. A cytosine was inserted between the 164th and 165th nucleotide from the termination codon. This mutation is a new mutation since it did not exist in patients of the control group and was not reported by other studies. Results of prediction showed that there were ten miRNAs which could bind to 3′UTR region of TBX1C gene and this mutation did not locate in binding regions of ten miRNAs and 3′UTR region of TBX1C gene. Conclusion A new mutation, c.*164_*165insC, exists in 3′UTR region of TBX1C gene. This mutation may affect the spatial configuration of binding of 3′UTR region of TBX1C gene and miRNAs and further affect the regulatory effect of miRNAs on the expression of TBX1C gene.

Key words: conotruncal heart defect, TBX1 gene, gene mutation, 3′UTR, microRNA