胚胎血红蛋白基因变异与早期自然流产的相关性研究

Correlation between the mutation of embryo hemoglobin genes and spontaneous abortion

目的:探讨胚胎血红蛋白基因变异与早期自然流产的相关性。方法:无菌收集40份早期自然流产的绒毛标本(自然流产组)及35份人工流产的绒毛标本(人工流产组),提取胚胎绒毛组织DNA,针对3个外显子及部分内含子、上游调控序列分片段设计引物,进行PCR扩增、测序,筛查基因突变。结果:所有胚胎绒毛组织血红蛋白ε、ζ基因的外显子片段均未发现突变。两组部分胚胎绒毛组织血红蛋白ε基因第一内含子84位点处发现4 bp CAAA插入序列,但其基因型频率比较差异无统计学意义(P>0.05)。两组部分胚胎绒毛组织血红蛋白ζ基因第一内含子128位点处发现58 bp插入序列,自然流产组血红蛋白ζ基因58 bp插入基因型频率高于对照组(P<0.05),关联性分析提示该基因片段变异与早期自然流产相关(r=0.26,P<0.05)。两组血红蛋白ζ基因58 bp插入纯合子基因型频率比较差异无统计学意义(P>0.05)。结论:胚胎基因血红蛋白ζ基因第一内含子第128位点处存在一58 bp插入变异,该变异与早期自然流产相关。

Objective： To investigate the association between the fetal hemoglobin （ε/ξ） genetic variants and early spontaneous abortion. Methods： Embryonic villi DNA was extracted from 40 embryo samples of early spontaneous abortion （spontaneous abortion group ） and 35 embryo samples of artificial abortion （artificial abortion group） ; then PCR-sequencing was carried out for detection of mutations for the three exons, some introns and the upstream regulatory sequence point fragments. Result： No mutation had been found in the all 3 extron segments in the screened genes of ε orξamong the villus tissues. A 4bp inserted sequence locating in the 58th locus was foundin the first intron of the ε gene, still no significant difference in this genotypic distribution between the two groups. However, the genotypic frequency of a 58bp inserted sequence found in the 128th locus in the first intron of the ξ gene was revealed with significantly higher frequency in spontaneous abortion group （P 〈 0. 05 ）. Furthermore, significant correlation was revealed between this mutation and spontaneous abortion （P 〈 0. 05 ）. Conclusions： The inserted sequence of 58bp in the first intron of the ξ gene is associated with early spontaneous abortion.