二代测序结合生物信息分析技术测定绝经后骨质疏松症病人血清外泌体来源相关miRNA的临床意义

徐靖泽; 朱成鑫; 陈绪鲲; 储佳奇; 邵松

doi:10.13898/j.cnki.issn.1000-2200.2023.12.011

二代测序结合生物信息分析技术测定绝经后骨质疏松症病人血清外泌体来源相关miRNA的临床意义

Clinical significance of identification of serum exosomal miRNA in postmenopausal osteoporosis patients by next-generation sequencing combined with bioinformatics analysis

摘要

摘要:
目的研究血清外泌体来源microRNA（miRNA）作为预测绝经期女性骨质疏松症的潜在生物标志物。
方法检测65岁以上骨质疏松症女性病人和健康对照者血清中的miRNA。RNA测序用于鉴定差异表达的miRNA。使用TargetScan、PicTar和miRanda数据库预测靶基因。通过STRING数据库构建蛋白质-蛋白质相互作用（protein protein interaction，PPI）网络，并使用Cytoscape软件进行可视化处理。最后，利用基因本体论（gene ontology，GO）和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）对目标靶基因的生物学功能进行分析。
结果绝经期女性骨质疏松病人中发现8个显著上调的miRNA。使用Cytoscope中的cytohubba插件，通过其网络功能计算出排名前十的核心基因为SOS1、ITGB3、SMAD4、PAK1、NRAS、PLCG1、MET、HGF、CCND1和CDKN1B。GO富集分析表明，靶基因主要集中在蛋白质结合、金属离子结合、DNA结合和ATP结合。PI3K-Akt信号通路和丝裂原活化蛋白激酶（MAPK）信号通路是最显著丰富的靶基因信号通路。
结论异常的血清外泌体miRNA与绝经期女性骨质疏松症有关，可作为预测绝经期女性骨质疏松症的潜在生物标志物。

Abstract:
ObjectiveTo study serum exosomal microRNA (miRNA) as potential biomarkers for predicting osteoporosis in postmenopausal women.
MethodsMiRNA in the serum of female patients with osteoporosis over 65 years old and healthy controls was detected.RNA sequencing was used to identify differentially expressed miRNA.TargetScan, PicTar, and miRanda databases were used to predict target genes.A protein protein interaction (PPI) network using the STRING database was constructed and visualized using Cytoscape software.Finally, the biological functions of the target gene were analyzed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG).
ResultsEight significantly upregulated miRNAs were found in postmenopausal female patients with osteoporosis.Using the cytohubba plugin in Cytoscope, the top ten core genes were calculated based on their network functionality, including SOS1, ITGB3, SMAD4, PAK1, NRAS, PLCG1, MET, HGF, CCND1, and CDKN1B.GO enrichment analysis showed that the target genes were mainly concentrated in protein binding, metal ion binding, DNA binding, and ATP binding.The PI3K-Akt signaling pathway and mitogen activated protein kinase (MAPK) signaling pathway were the most significantly enriched target gene signaling pathways.
ConclusionsAbnormal serum exosomal microRNAs are associated with postmenopausal osteoporosis and can serve as potential biomarkers for predicting osteoporosis in postmenopausal women.

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