CAJ | 학술논문

目的通过对低密度脂蛋白受体相关蛋白5/6 (low density lipoprotein receptoPrelated protein 5/6,LRPS/6)基因单核苷酸多态性(single nucleotide polymorphisms,SNPs)基因型与先天性巨结肠症(Hirschsprung disease's,HD)的相关性研究,探索可能潜在的致病基因.方法采集200例临床确诊的HD患儿(病例组),200名健康儿童(对照组)的外周静脉血,提取白细胞基因组DNA,PCR扩增LRP5/6基因4个位点(rs121908674,rs121908671/rs121918313,rs117554756),将PCR产物测序进行突变筛选,明确突变位点和类型,并进一步结合病例对照和生物信息学分析探讨基因变异的意义.结果病例组与对照组LRP5 rs121908671、LRP6 rs117554756AA和AG、AA和AC基因型频率无显著性差异(P＞0.05);而病例组与对照组LRP5 rs121908674 CC、CG和GG基因型频率及C和G等位基因频率差异显著(P＜0.05),CC和CG基因型及G等位基因的患病风险分别为0.005、0.195和0.001;LRP6 rs121918313 CC、CT和TT基因型频率及C和T等位基因频率差异显著(P＜0.05),CC和CT基因型及T等位基因的患病风险分别为0.012、0.504和0.003.LRP6基因rs121918313测序检出有杂合性缺失和单碱基的替换,第131位密码子核苷酸AAA→CAA.结论 LRP5/6基因多态性位点可能对HD有影响,并且可能有一定的遗传性.
목적 통과대저밀도지단백수체상관단백5/6 (low density lipoprotein receptoPrelated protein 5/6,LRPS/6)기인단핵감산다태성(single nucleotide polymorphisms,SNPs)기인형여선천성거결장증(Hirschsprung disease's,HD)적상관성연구,탐색가능잠재적치병기인.방법 채집200례림상학진적HD환인(병례조),200명건강인동(대조조)적외주정맥혈,제취백세포기인조DNA,PCR확증LRP5/6기인4개위점(rs121908674,rs121908671/rs121918313,rs117554756),장PCR산물측서진행돌변사선,명학돌변위점화류형,병진일보결합병례대조화생물신식학분석탐토기인변이적의의.결과 병례조여대조조LRP5 rs121908671、LRP6 rs117554756AA화AG、AA화AC기인형빈솔무현저성차이(P＞0.05);이병례조여대조조LRP5 rs121908674 CC、CG화GG기인형빈솔급C화G등위기인빈솔차이현저(P＜0.05),CC화CG기인형급G등위기인적환병풍험분별위0.005、0.195화0.001;LRP6 rs121918313 CC、CT화TT기인형빈솔급C화T등위기인빈솔차이현저(P＜0.05),CC화CT기인형급T등위기인적환병풍험분별위0.012、0.504화0.003.LRP6기인rs121918313측서검출유잡합성결실화단감기적체환,제131위밀마자핵감산AAA→CAA.결론 LRP5/6기인다태성위점가능대HD유영향,병차가능유일정적유전성.
Objective To explore the relationship between low density lipoprotein receptorre lated protein 5/6 (LRPS/6) genes and the genotypes of single nucleotide polymorphisms (SNPs) of Hirschsprung's disease (HD).Methods A total of 200 children of HD (case group) and 200 healthy children (control group) were recruited to obtain genomic DNA from peripheral blood leukocytes.Pol ymerase chain reaction (PCR) amplification was performed for LRPS/6 genes (rs121908674,rs121908671/rs121918313,rs117554756) and PCR products were sequenced for mutation screening.Cas-control study and bioinformatic analyses were utilized to explore the potential roles of variations.Results No significant inter group differences existed in allelic and genotypic frequencies of AA and AG for rs121908671 (P＞0.05).LRP6 gene rs117554756 allelic and genotypic frequencies of AA and AC existed between patients with HD and the control group (P＞0.05).The allelic and genotypic fre quencies in LRP5 gene rs121908674 as well as the genotypes of CC,CG and GG were related in two groups (P＜0.05).C and G polymorphisms were present in HD.The disease risks of genotype of CC and GG and allelic gene of G were 0.005,0.195 and 0.001 respectively.The allelic and genotypic fre quencies in LRP6 gene rs121918313 and the genotypes of CC,CT and TT were related in two groups (P＜0.05).C and T polymorphisms were present in HD.The disease risks of genotype of CC and CT and allelic gene of T were 0.012,0.504 and 0.003 respectively.And rs121918313 sequencing demon strated a loss of heterozygosity and single nucleotide substitution.The single nucleotide substitution for rs121918313 was confirmed in HD:AAA→CAA at position 131.Conclusions The polymor phisms may affect HD in LRP5/6 genes so as to carry a hereditary tendency.