中国新疆地区哈萨克族和汉族扩张型心肌病患者TPM1基因的筛查

TPM1 gene mutation is associated with dilated cardiomyopathy in Kazaks in Xinjiang

目的 探讨中国新疆地区哈萨克族和汉族扩张型心肌病患者TPM1基因的突变特点,并明确二者的关联性.方法 入选一哈萨克族扩张型心肌病家系,成员共31名,特发性扩张型心肌病（IDCM）患者100例（哈萨克族50例、汉族50例）,以及100例健康志愿者（哈萨克族50例、汉族50例）,对入选者行TPM1基因筛查.样本均为2012年至2014年新疆医科大学第一附属医院门诊及住院患者.提取入选者外周血DNA,聚合酶链反应（PCR）扩增TPM1基因的9个外显子和外显子附近的内含子,对目的基因进行正反方向测序,测序结果与CHROMAS软件和Pubmed中BLAST软件的标准模板进行对比,并分析突变位点与哈萨克族IDCM和健康志愿者、汉族IDCM和健康志愿者之间的关系.采用酶联免疫吸附试验（ELISA）对入选者原肌球蛋白进行定性、定量检测.结果 在哈萨克族家族性扩张型心肌病的2名成员中检测到TPM1基因3号外显子一未报的单个碱基变异c.524 G＞T,导致谷氨酰胺转变为组氨酸（Gln1 11 His）.哈萨克族IDCM患者TPM1基因多态位点rs1071646的基因型分布和等位基因频率与健康志愿者比较差异均有统计学意义（分别为x2=13.36、P=0.001和x2=10.25、P=0.001）,而汉族IDCM患者与健康志愿者二者比较差异则均无统计学意义（P均＞0.05）.IDCM患者（包含哈萨克族和汉族）的原肌球蛋白含量低于健康志愿者（包括哈萨克族和汉族）[（1 764.2±350.9） ng/L比（2 369.7±345.9） ng/L,P=0.001].结论 新疆哈萨克族和汉族的IDCM患者中TPM1基因多态位点rs1071646发生率均较高.TPM1基因多态位点rs1071646可能是新疆哈萨克族人群发生扩张型心肌病的一个易感基因多态位点.

Objective Detect the relationship between TPM1 gene mutations and dilated cardiomyopathy （DCM） of Kazaks and Hans in Xinjiang.Methods TPM1 gene was screened from 31 family members in a Kazak family with familiar DCM （FDCM）,100 patients with idiopathic DCM （IDCM,50 Kazaks and 50 Hans）,and in 100 healthy controls （50 Kazaks and 50 Hans）.All the samples were the inpatients or outpatients of First Affiliated Hospital of Xinjiang University from 2012 to 2014.PCR was used to amplify 9 exons and nearby introns of the TPM1 gene.The amplified products were sequenced and compared with the standard sequence with CHROMAS software and BLAST software in Pubmed to identify mutation sites.The relationship between TPM1 gene mutations in the Kazak IDCM and healthy volunteers,between Han and Kazak IDCM and healthy volunteers was analyzed.Tropomyosin was qualitatively and quantitatively detected by ELISA in all subjects.Results A novel variant （c.524 G 〉 T） was identified in two FDCM patients at exon 3,this mutation caused an amino acid substitution,Gln11 1His.The FDCM,IDCM from Kazak and Han,healthy volunteers from Kazak and Han were founded a rs1071646 （c.644C 〉A,Ala1S1Ala）.There was a significant difference in the genotype distribution （x2 =13.36,P =0.001） and allele frequency （x2 =10.25,P =0.001） between Kazaks with IDCM and Kazak controls of rs1071646,while these parameters were similar between Han IDCM patients and Han controls （all P 〉 0.05）.The tropomyosin content of Kazak and Han IDCM patients were significantly lower than Kazak and Han controls （（1 764.2 ± 350.9） ng/L vs.（2 369.7 ± 345.9） ng/L,P =0.001）.Conclusion TPM1 gene of rs1071646 polymorphism is a possible independent risk factor for IDCM in Kazaks but not Han Chinese.