Association Of Mismatch Repair Protein Gene Polymorphism With Oligospermia And Azoospermia

Objective:To establish the methodology of tetra-primer amplification refractory polymorphism system-PCR(4P-ARMS-PCR).Analyze the association of mismatch repair protein gene polymorphism with oligospermia and azoospermia,in order to explore new ways of studying the etiology of oligospermia and azoospermia.Methods:1.Using mismatch repair protein MLH3 gene C2531T polymorphism as an example to establish the methodology of 4P-ARMS-PCR.2.Detecting mismatch repair protein MSH4 gene G289A polymorphism by 4P-ARMS-PCR.3.Detecting mismatch repair protein MSH4 gene A1766G polymorphism by 4P-ARMS-PCR.4.Detecting mismatch repair protein MSH5 gene C85T polymorphism by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).Results:1.PCR-RFLP results,4P-ARMS-PCR results and sequence results of detecting mismatch repair protein MLH3 gene C2531T polymorphism were fully consistent.2.The distribution of genotypes(MLH3 C2531T) in normal controls and oligospermia,azoospermia patients did not differ significantly from expected under Hardy-Weinberg equilibrium(oligospermia, azoospermia patients:x~2=0.012,P＞0.05;normal controls:x~2=1.430, P＞0.05).That was to say the selected samples were representative. The genotype distribution in patients and controls was significantly different,and there was a higher prevalence of CT+TT genotypes in patients(x~2=8.695,P＜0.05,OR=1.975,95%CI:1.230～3.173);The T allele frequency was significantly higher in patients than in controls (x~2=10.631,P＜0.05,OR=2.250,95%CI:1.352～3.744).3.Not able to find mismatch repair protein MSH4 gene G289A polymorphism.4.Not able to find mismatch repair protein MSH4 gene A1766G polymorphism.5.The distribution of genotypes(MSH5 C85T) in normal controls and oligospermia,azoospermia patients did not differ significantly from expected under Hardy-Weinberg equilibrium(oligospermia, azoospermia patients:x~2=0.113,P＞0.05;normal controls:x~2=0.626, P＞0.05).That was to say the selected samples were representative. The genotype distribution in patients and controls was significantly different,and there was a higher prevalence of CT+TT genotypes in patients(x~2=11.829,P＜0.05,OR=2.507,95%CI:1.428～4.400);The T allele frequency was significantly higher in patients than in controls (x~2=14.195,P＜0.05,OR=2.887,95%CI:1.599～5.211).6.The joint analysis of the genotypes of MLH3 C2531T and MSH5 C85T showed that:the genotype distribution in patients and controls was significantly different(x~2=18.181,P＜0.05);Single person expressing MLH3 C2531T(CT+TT) and MSH5 C85T(CT+TT) genotypes at the same time has the greatest risk of oligospermia,azoospermia morbidity(x~2=15.653,P=0.000,OR=6.775,95%CI:2.117～21.683).Conclusion:1.4P-ARMS-PCR can be a rapid,simple and accurate detection of gene polymorphism of single nucleotide polymorphism(SNP). 2.Mismatch repair protein MLH3 gene C2531T polymorphism was associated with oligospermia and azoospermia,CT+TT genotype maight be a risk factor of disease,and C allele maight be a genetic susceptibility gene of the disease.3.Mismatch repair protein MSH5 gene C85T polymorphism was associated with oligospermia and azoospermia,CT+TT genotype maight be a risk factor of disease,and C allele maight be a genetic susceptibility gene of the disease.4.Mismatch repair proteins MLH3 and MSH5 maight have some intrinsic relevance.The polymorphisms of MLH3 C2531T and MSH5 C85T interacting with each other might result in the occurrence of oligospermia and azoospermia.