CAJ | 학술논문

目的研究先天性巨结肠与RET基因多态性的关系,进一步推断主要的先天性巨结肠相关单倍型,并分析浙江汉族人群RET基因单核苷酸多态性的分布特点.方法采集2005年至2007年浙江大学医学院附属儿童医院收治的123例汉族先天性巨结肠患儿以及194例健康体检儿童的外周血,并提取DNA.根据选取的RET基因单核苷酸多态性合成引物,行PCR扩增,经2%琼脂糖电泳证实PCR产物后,将PCR产物进一步纯化、测序.采用PHASE软件计算单倍型频率.将健康儿童RET基因各位点等位基因频率与文献和数据库已有的其他种族资料进行比较.RET基因和先天性巨结肠间关系的分析采用x2检验,以比值比及95%可信区间表示.结果 RET基因-5G＞A、-1A＞C、c135G＞A、c2307T＞G 4个位点的少见基因型AA、CC、AA、GG在先天性巨结肠患儿中的频率显著高于健康儿童(x2=57.775,20.469,57.040,38.869,P＜0.05).RET基因-5A、-1C、c135A、c2307G在先天性巨结肠患儿中等位基因频率显著高于健康儿童(x2=85.114,53.117,77.005,70.161,P＜0.05).RET基因各位点等位基因频率在不同类型先天性巨结肠间的差异无统计学意义(x2=0.048,0.265,0.395,0.027,P＞0.05).RET基因4个位点单倍型中ACAG占先天性巨结肠患儿的75.2%,显著高于健康儿童的38.7%(x2=62.776,P＜0.05).本组健康儿童RET基因-5A、c135A和c2307G位点的少见等位基因频率明显高于欧洲高加索人和非洲约鲁巴人(P＜0.05).结论 RET基因-5G＞A、-1A＞C、c135G＞A、c2307T＞G 4个位点多态性与浙江汉族人群先天性巨结肠发病明显相关,但与先天性巨结肠类型无关.RET基因4个位点单倍型中ACAG是浙江汉族人群的先天性巨结肠相关核心单倍型.浙江汉族人群的RET基因-5、c135和c2307位点的少见等位基因频率显著高于欧洲高加索人和非洲约鲁巴人.
목적 연구선천성거결장여RET기인다태성적관계,진일보추단주요적선천성거결장상관단배형,병분석절강한족인군RET기인단핵감산다태성적분포특점.방법 채집2005년지2007년절강대학의학원부속인동의원수치적123례한족선천성거결장환인이급194례건강체검인동적외주혈,병제취DNA.근거선취적RET기인단핵감산다태성합성인물,행PCR확증,경2%경지당전영증실PCR산물후,장PCR산물진일보순화、측서.채용PHASE연건계산단배형빈솔.장건강인동RET기인각위점등위기인빈솔여문헌화수거고이유적기타충족자료진행비교.RET기인화선천성거결장간관계적분석채용x2검험,이비치비급95%가신구간표시.결과 RET기인-5G＞A、-1A＞C、c135G＞A、c2307T＞G 4개위점적소견기인형AA、CC、AA、GG재선천성거결장환인중적빈솔현저고우건강인동(x2=57.775,20.469,57.040,38.869,P＜0.05).RET기인-5A、-1C、c135A、c2307G재선천성거결장환인중등위기인빈솔현저고우건강인동(x2=85.114,53.117,77.005,70.161,P＜0.05).RET기인각위점등위기인빈솔재불동류형선천성거결장간적차이무통계학의의(x2=0.048,0.265,0.395,0.027,P＞0.05).RET기인4개위점단배형중ACAG점선천성거결장환인적75.2%,현저고우건강인동적38.7%(x2=62.776,P＜0.05).본조건강인동RET기인-5A、c135A화c2307G위점적소견등위기인빈솔명현고우구주고가색인화비주약로파인(P＜0.05).결론 RET기인-5G＞A、-1A＞C、c135G＞A、c2307T＞G 4개위점다태성여절강한족인군선천성거결장발병명현상관,단여선천성거결장류형무관.RET기인4개위점단배형중ACAG시절강한족인군적선천성거결장상관핵심단배형.절강한족인군적RET기인-5、c135화c2307위점적소견등위기인빈솔현저고우구주고가색인화비주약로파인.
Objective To study the relationship between polymorphisms of RET gene and the incidence of Hirschsprung's disease, investigate the haplotypes of RET gene in patients with Hirschsprung's disease, and analyze the characteristics of single nucleotide polymorphisms of RET gene in Zhejiang Han population. Methods Peripheral blood samples were collected from 123 patients with Hirschsprung's disease and 194 healthy children from 2005 to 2007 at Children's Hospital of Zhejiang University. G enomic DNA was extracted from the peripheral blood, and the genotypes were analyzed using polymerase chain reaction and direct sequencing. The frequencies of haplotypes were estimated using the PHASE software. The frequencies of RET alleles of the 194 healthy children were compared with those of other races. The correlation between RET gene and Hirschsprung's disease was analyzed using the chi-square test, and it was expressed in the form of odds ratio and 95% confidence interval.Results Increased risk of Hirschsprung's disease was observed in homozygous genotypes of the RET alleles -5AA, - 1CC, c135AA and c2307GG when compared with other genotypes ( x2 =57. 775, 20.469, 57. 040,38. 869, P ＜ 0. 05 ). Increased risk of Hirschsprung's disease was also observed in RET alleles -5A, - 1 C,c135A, c2307G when compared with other alleles ( x2 = 85.114, 53.117, 77. 005, 70. 161, P ＜0.05). There was no relationship between the frequencies of the alleles and the types of Hirschsprung's disease ( x2= 0.048,0.265, 0. 395, 0.027, P ＞ 0.05 ). The percentage of patients with haplotype ACAG was 75.2%, which was significantly higher than that of 38.7% in healthy children ( x2 = 62. 776, P ＜ 0.05 ). The frequencies of Hirschsprung's disease-associated alleles of RET - 5A, c135A and c2307G in Chinese were significantly higher than thosein the Caucasians or Yorubas (P＜0.05). Conclusions RET -5G＞A, -1A ＞C, c135G＞A and c2307T ＞ G are associated with Hirschsprung's disease and haplotype ACAG is the core one in Zhejiang Han population. There is no relationship between the frequency of allele and the types of Hirschsprung's disease. The frequencies of alleles of RET -5A, c135A and c2307G in Chinese are significantly higher than those in the Caucasians or Yorubas.