中文摘要：

目的 探讨HIRA基因3＇UTR区与法洛四联症的关系及相关微RNA.方法 病例组为于2007年4月至2012年12月复旦大学附属儿科医院通过心导管检查及外科手术证实为法洛四联症的患儿,共278例;对照组为同期本院门诊体检儿童,共515名.对其进行HIRA基因3＇UTR区DNA序列分析,用生物信息学预测可能与其结合的微RNA,通过双荧光素酶报告基因检测系统及实时定量PCR验证微RNA对目的基因的调控作用.对两组结果通过x2检验或t检验进行比较.结果 病例组和对照组中HIRA基因3＇UTR区rs：117447448单核苷酸多态性（SNP）位点的分布差异有统计学意义[GA和（或）AA基因型分别为11.5％（32/278）和4.9％（25/515）,P=0.001].在线预测rs：117447448 SNP位点上游10 bp存在微RNA328的结合位点.miR328在心脏表达,且与心肌梗死与心房颤动相关.共同转染p-silencer＋ miR328质粒和pgl3-promoter＋ HIRA3＇UTR质粒293细胞荧光素酶的活性低于单独转染pgl3-promoter＋ HIRA3＇UTR质粒的活性（0.012 5±0.000 6比0.019 6±0.003 8,P=0.034）.转染p-silencer＋ miR328质粒的293T细胞HIRA基因的表达较转染p-silencer空载质粒的表达低（1.039 6±0.077 2比1.608 7±0.274 9,P=0.037）.用rs：117447448SNP位点不同基因型的质粒与miR328共同转染293细胞,不同基因型的荧光素酶活性差异无统计学意义（P=0.380）.结论 HIRA基因3＇UTR区rs：117447448SNP位点可能与法洛四联症的发生相关;HIRA基因是微RNA328的靶基因.rs：117447448SNP位点不是微RNA328作用于HIRA基因的主要靶位.

英文摘要：

Objective To explore the HIRA gene sequences of 3＇UTR region and elucidate the role of 3＇UTR region of HIRA gene in the pathogenesis of tetralogy of Fallot （TOF）.Methods Patients of TOF were confirmed by cardiac catheterization or surgery between April 2007 and December 2012 at our hospital.Mutations and single nucleotide polymorphisms （SNPs） were screened in 278 unrelated probands with isolated TOF and 515 controls.Target Scan was used to predict micro RNAs with possible combinations with 3 ＇UTR region of HIRA gene.Dual-luciferase assay and real-time PCR were performed to detect the inhibition activity of micro RNAs on target genes.And x2 and t tests were used to analyze the results.Results Statistically significant change occurred in the alleleic frequencies of existing SNPs （rs：117447448） between TOF patients and control group （11.5％ （32/278） vs 4.9％ （25/515）,P =0.001）.The combining site of miR328 was predicted to be 10 bp upstream of SNP site.MiR328 was expressed in heart and it was related with myocardial infarction and atrial fibrillation.Dual-luciferase assay showed a decreased level of luciferase after co-transfection with miR328 （0.012 5 ±0.000 6 vs 0.019 6 ±0.003 8,P =0.034）.So was the expression of HIRA （1.039 6 ±0.077 2 vs 1.608 7 ±0.274 9,P =0.037）.However,the luciferase level was not affected by SNP （rs：117447448） （P=0.380）.Conclusions The SNP （rs：117447448） of 3＇UTR region of HIRA gene is related with TOF.HIRA is the target gene of miR328.Although SNP （rs：117447448） is not a major site of target gene HIRA for micro RNA328,it provides an important clue to indepth studies of 3＇UTR region of HIRA gene in the pathogenesis of TOF.