PLCG1、KIF2B和KIF4B基因与完全受精失败的相关性分析

Correlation analysis of PLCG1,KIF2B and KIF4B genes with complete fertilization failure

目的筛选与完全受精失败相关的基因,探讨完全受精失败的分子机制。方法对64对IVF完全受精失败夫妇(实验组)和157对IVF受精正常夫妇(对照组)进行全外显子测序,AdapterRemoval软件对测序结果进行处理,采用Sentieon软件的Haplotyper算法进行单核苷酸多态性(SNP)检测,根据不同分组SNP位点的突变频率,采用R语言chisq.test(data)命令进行卡方检验(chi-square test),分析找到完全受精失败相关的候选基因,使用公共数据库对候选基因的功能进行分析。结果在女性和男性群体中分别获得148和120个与完全受精失败相关的候选基因,分别含有164和132个突变位点。经基因显著性分析及数据库功能验证,在女性群体中筛选到与完全受精失败相关的PLCG1基因(rs2228246),实验组的突变率[6.15%(4/64)]显著高于对照组(0/157)(P<0.05)。在男性群体中发现与完全受精失败相关的KIF2B(rs76337756)和KIF4B(rs10056252)基因,实验组中突变率为4.61%(3/64),显著高于对照组(0/157)(P<0.05)。结论在IVF完全受精失败夫妇中筛选到PLCG1、KIF2B和KIF4B基因,这为后期探究完全受精失败过程中的分子机制奠定基础。

Objective:To screen the genes related to complete fertilization failure and explore the molecular mechanism of complete fertilization failure.Methods:The whole exome sequencing was performed on 64 IVF complete fertilization failure couples(experimental group)and 157 normal IVF fertilization couples(control group).The sequencing results were processed by AdapterRemoval software.The Haplotyper algorithm of Sentieon software was used for SNP detection.According to the mutation frequency of SNP loci in different groups,using R language chisq.test(data)command to perform chi-square test to analyze and find the candidate genes related to complete fertilization failure.The function of candidate genes was analyzed using a public database.Results:A total of 148 and 120 complete fertilization failure-related candidate genes,containing 164 and 132 mutation sites,were found in female and male group respectively.After gene dominance analysis and database function verification,PLCG1 gene(rs2228246)related to complete fertilization failure was screened in female group.The mutation rate in the experimental group[6.15%(4/64)]was significantly higher than that in the control group(0/157)(P<0.05).KIF2B(rs76337756)gene and KIF4B(rs10056252)gene related to TFF were found in male group.The mutation rate in the experimental group was 4.61%(3/64),which was significantly higher than that in the control group(0/157)(P<0.05).Conclusions:PLCG1,KIF2B and KIF4B genes were screened from the couples with complete fertilization failure,which laid the foundation for exploring the molecular mechanism of complete fertilization failure in the later stage.