弱精子症患者精子lncRNA与mRNA基因表达特征分析

Gene expression characteristics of lncRNAs and mRNAs in the sperm of asthenospermia patients

目的:通过DNBSEQ平台测序技术构建文库,并以此来确定人类正常与弱精子症精子中lncRNA和mRNA的表达差异,并通过生物信息学方法分析其在弱精子症中的生物学意义。方法:将9份正常精液样本及9份弱精子症精液样本分组后,分离精子,提取总RNA,通过DNBSEQ测序平台检测得到RNA-seq在精子中的表达情况,通过GO富集分析和KEGG通路分析,进一步分析其相关功能情况。结果:使用DNBSEQ平台检测,每组平均产出10.64G数据,共检测到282185个RNA,包含有107009个lncRNA,其中15157个lncRNA具有表达差异,2190个lncRNA表达上调,12967个表达下调;共检测到19514个mRNA,其中13736个mRNA具有表达差异,4995个mRNA表达上调,8741个mRNA表达下调。差异基因主要富集在精子细胞膜、离子通道的功能以及精子发育、受精相关的通路上。结论:通过对弱精子症与正常精子的测序分析,鉴别出差异表达的lncRNA和mRNA,可通过胞膜离子通道对精子功能进行调控,进而导致弱精子症的发生,可为弱精子症的进一步研究提供分子基础。

Objective:To determine the differential expressions of long non-coding RNA(lncRNA)and messenger RNA(mRNA)in normal and asthenospermia(AS)men and analyze their biological significance in AS.Methods:We isolated and extracted total RNAs from 9 normal and 9 AS sperm samples,determined the expressions of RNAs in the sperm using the DNBSEQ sequencing platform,and analyzed their relevant functions by gene ontology enrichment(GO)and Kyoto encyclopedia of genes and genomes pathway(KEGG)analyses.Results:An average of 10.64G data was generated per group,with 282185 RNAs detected,including 107009 lncRNAs.Among the total number of lncRNAs,15157 were differentially expressed,2190 upregulated and 12967 downregulated;and among the 19514 mRNAs,13736 were differentially expressed,4995 upregulated and 8741 downregulated.Differentially expressed genes were enriched mainly in the sperm cell membrane and the pathways related to the ion channel functions,sperm development and fertilization.Conclusion:Differentially expressed lncRNAs and mRNAs can be identified by sequencing analysis of AS and normal sperm.Regulation of sperm function through membrane ion channels may contribute to the development of AS,which provides a molecular basis for further research on AS.