mtATPase6基因变异与弱精子症的相关分析

Mutation Analysis of the mtATPase6 Associated with Asthenospermia Patients

为了分析mtATPase6基因突变与弱精子症的相关性,按WHO标准收集了27例弱精子症精液标本和28例精子活力正常精液标本,PCR扩增mtATPase6基因,纯化测序,分析mtATPase6基因突变,比较两组突变频率的差异。结合生物信息学工具分析错义突变位点的氨基酸进化保守性及其蛋白质部分三级结构。结果显示:发现了6个未曾报道过的突变位点;弱精子症组mtATPase6基因平均突变率显著高于对照组,可能与弱精子症有一定的相关性。G8584A、A8701G和G9053A三个错义突变可能是多态性位点,其余8个错义突变中的6个具有进化保守性的位点累计突变频率显著高于对照组,这些位点突变可能与弱精子症有关。

This research attempted to investigate whether there was correlationship between mtATPase6 point mutation in sperms and asthenospermia. After 27 asthenospermia cases and 28 control cases were collected under the WHO criterion, the regions of nt 7908-8816 and 8602-9416 of them were amplified by using PCR of two pairs primers. Consequently, the point mutation, mutation rate as well as evolutionary conservation pattern were analyzed by employing sequencing technology and bioinformatics tools. As a result, 20 point mutations were found, among which six had never been identified before： A8586G, C8811T, T8825C, T8966C, C9071T and A9120G. Meanwhile, a significant difference （P〈O.05） was detected in the average mutation rate between asthenospermia （1.52±0.98） and control groups （0.96±0.84）, which suggests that a remarkable increase of mtATPase6 average mutation rate may correlate with asthenospermia in asthenospermia group. The three specific missense point mutation, namely, the G8584A, A8701G and G9053A, may be potential polymorphism. Besides, the differences between asthenospermia （22.22%） and control groups （3.57%） were statistically significant （P〈0.05） in terms of the cumulative mutation rates for the remaining six sites with evolution conservation, which may have the relationship with asthenospermia.