AXIN2基因三个位点与先天性巨结肠的关联研究

目的 探讨AXIN2基因单核苷酸多态性(single nucleotide polymorphisms,SNPs)rs2240308、rs8081536和rs9913621 3个位点与先天性巨结肠(Hirschspnmg disease,HSCR)的关联性.方法 对120例HSCR患者和120名正常人群外周血进行基因组DNA抽提,用PCR技术对AXe2基因3个位点(rs2240308、rs8081536和rs9913621)进行PCR扩增,PCR产物用内切酶CviJ I、Dde I和BaN I消化,将SNPs位点进行分型与分析,应用X2检验统计分析病例组和对照组等位基因频率、等位基因型频率及其患病风险;同时将PCR产物进行测序,以进一步确定基因突变位点.结果 HSCR组与对照组AXIN2 rs8081536 CC和CT基因型频率差异无统计学意义(P＞0.05);而HSCR组与对照组AXIN2 rs2240308 GG、AG和从基因型频率及A和G等位基因频率差异有统计学意义(P＜0.05),GC和从基因型及G等位基因的患病风险分别为2.091、0.846和1.703;HSCR组与对照组AXIN2 rs9913621 CC、CT和TT基因型频率及C和T等位基因频率差异有统计学意义(P＜0.05),CC和TT基因型及T等位基因的患病风险分别为0.535、1.113和1.569.测序rs2240308第301位密码子核苷酸GCA→CCA杂合突变;rs913621第199位密码子核苷酸CAC→CAG杂合突变.结论 AXIN2 rs8081536等位基因变异与HSCR的易感性无关;AXIN2 rs2240308和rs9913621与HSCR的发生可能有关联,具有GG基因型与CC基因型患HSCR的危险性相对较高.

Association of single nucleotide polymorphisms of Axis inhl'bitor-2 gene rs224030,rs8081536,rs9913621 with Hirschsprung disease

Objective To investigate the association of Axis inhihitor-2 (AXIN2) gene rs2240308,rs8081536 and rs9913621 single nucleotide polymorphisms (SNPs) with Hirschsprung disease(HSCR).Methods DNA was ex tracted from 120 HSCR patients and 120 healthy controls.The AXIN2 gene exon2-rs2240308,exon5-rs8081536 and exon6-rs9913621 were amplified by polymerase chain reaction (PCR).SNPs of AXIN2 gene were analyzed by restrictive endonuclease digestion with CviJ I,Dde I and BstN I and DNA sequencing.The allele and genotype frequencies and risk factors of HSCR and control group were analyzed by Chi-square test.Results No significant differences were found in genotype frequencies of CC and CT in AXIN2 rs8081536 between HSCR patients and the control group (P＞0.05).The frequencies of genotypes GG,AG and AA as well as alleles A and G genotypes in AXIN2 gene rs2240308 locus were found to be associated with HSCR (P＜0.05).The disease risk of genotypes GG and AA and allele G with was 2.091,0.846 and 1.703,respectively.The frequencies of genotypes CC,CT and TT as well as alleles C and T in AXIN2 gene rs9913621 locus were also associated with HSCR (P＜0.05).The disease risk of the genotypes CC and TT and the allele T was 0.535,1.113 and 1.569,respectively.Heterozygote mutation for rs2240308 was found in the HSCR patients,i.e.the GCA→CCA mutation at position 301.Heterozygosity for rs9913621 was observed in the HSCR patients,i.e.the CAC→CAG mutation at position 199.Conclusion The rs8081536 allelic variation in AXIN2 gene does not contribute to the susceptibility of HSCR in the patients.AXIN2 rs2240308 and rs9913621 allelic variation might be related to HSCR.Individuals having allele G and T in these loci are at relatively high risk for HSCR.