hsa-miR-21-5p靶基因预测及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2023.11.004

摘要/Abstract

摘要： 【目的】试验旨在对hsa-miR-21-5p的序列保守性和靶基因进行生物信息学分析，为进一步探讨其在卵母细胞成熟过程中的功能及调节机制提供思路。【方法】从GEO数据库获取人卵母细胞不同成熟阶段对应的卵丘细胞基因表达谱数据（GSE31681），利用R软件筛选差异表达基因（DEGs），通过FunRich软件预测DEGs的靶向miRNAs，找出hsa-miR-21-5p靶向的DEGs，获得miRNA-mRNA关系对；利用BLAST程序进行相似性比对，通过Mega 11.0软件构建系统进化树，采用ViennaRNA Web Services预测pre-miR-21-5p的二级结构；使用miRTarBase网站预测hsa-miR-21-5p靶基因，并进行GO功能和KEGG通路富集分析；通过STRING数据库建立蛋白质互作网络，并利用Cytoscape软件进行可视化分析。【结果】共筛选出7个卵母细胞成熟过程中GV/MⅡ期对应卵丘细胞中hsa-miR-21-5p靶向的DEGs。对物种间miR-21-5p序列分析发现，人pre-miR-21-5p与小鼠、褐家鼠、黄牛、绵羊、笔尾树鼩、猕猴的相似性均达95%以上，且miR-21-5p在进化过程中高度保守。ViennaRNA Web Services预测结果显示，pre-miR-21-5p的二级结构具有典型的单一茎环结构。靶基因预测发现，hsa-miR-21-5p共有潜在的723个靶基因。GO功能富集结果显示，靶基因主要富集到细胞增殖调控、内源性凋亡信号通路、蛋白激酶活性的正向调节等功能。KEGG通路富集表明，靶基因主要富集到MAPK和p53等信号通路。蛋白互作分析发现，靶基因MYC、FOXO3、STAT3等编码的蛋白与其他蛋白存在较多靶向关系，参与了PI3K/Akt、JAK-STAT等信号通路。【结论】miR-21-5p在生物进化过程中具有高度保守性，其主要通过调节卵丘细胞中相关基因的表达来调控MAPK和p53等信号通路，进而在卵母细胞成熟过程中发挥作用。

关键词: miR-21-5p; 靶基因; 卵丘细胞; 生物信息学分析

Abstract: 【Objective】 This study was aimed to analyze the sequence conservation and target genes of hsa-miR-21-5p by bioinformatics, so as to provide ideas for further exploring its function and regulatory mechanism during oocyte maturation.【Method】 The gene expression profiles (GSE31681) of cumulus cell at different stages of human oocytes during maturation were obtained from GEO database.The differentially expressed genes (DEGs) were screened by R software.The miRNAs targeted with DEGs were predicted by FunRich software and found the DEGs targeting by hsa-miR-21-5p to obtain miRNA-mRNA relationship pairs.The similarity and phylogenetic tree were analyzed and constructed by BLAST program and Mega 11.0 software, respectively, and the secondary structure of pre-miR-21-5p were predicted by ViennaRNA Web Services.The target genes of hsa-miR-21-5p were predicted using miRTarBase website, and GO function and KEGG pathway enrichment analysis was carried out.The STRING online database was accessed to develop the protein interaction network and the result was visualized through Cytoscape software.【Result】 A total of 7 DEGs targeted by hsa-miR-21-5p in cumulus cells corresponding to GV/MⅡ stages during oocyte maturation were screened.The sequence analysis of miR-21-5p among species showed that thesimilarity of human pre-miR-21-5p with Mus musculus, Rattus norvegicus, Bos taurus, Ovis aries, Tupaia chinensis and Macaca mulatta was over 95%, and miR-21-5p was highly conserved during evolution.The ViennaRNA Web Services prediction results showed that the secondary structure of pre-miR-21-5p had a typical single stem-loop structure.Target genes prediction revealed that hsa-miR-21-5p had 723 potential target genes.GO function analysis results showed that the target genes were mainly enriched in regulation of cell proliferation, intrinsic apoptotic signaling pathway, positive regulation of protein kinase activity and so on.KEGG pathway analysis results showed that the target genes were mainly enriched in MAPK and p53 signaling pathways.Protein interaction analysis results revealed that the proteins encoded by the target genes MYC, FOXO3 and STAT3 had more targeted relationships with other proteins, and involved in PI3K/Akt, JAK-STAT and other signaling pathways.【Conclusion】 miR-21-5p was highly conserved in biological evolution and played a role in oocyte maturation mainly by regulating the expression of related genes to regulate signaling pathways such as MAPK and p53 in cumulus cells.

Key words: miR-21-5p; target gene; cumulus cells; bioinformatics analysis

中图分类号:

Q78
S814.8

张昕沛, 吴家乐, 吴飞, 狄生伟. hsa-miR-21-5p靶基因预测及生物信息学分析[J]. 中国畜牧兽医, 2023, 50(11): 4349-4358.

ZHANG Xinpei, WU Jiale, WU Fei, DI Shengwei. Target Gene Prediction and Bioinformatics Analysis of hsa-miR-21-5p[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(11): 4349-4358.

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