多囊卵巢综合征患者颗粒细胞miRNA及mRNA表达谱的构建

Construction of microRNA and mRNA expression profiles in granulosa cells of patients with polycystic ovary syndrome

目的构建多囊卵巢综合征(polycystic ovary syndrome,PCOS)患者卵巢颗粒细胞中的微小RNA(miRNA)和mRNA表达谱。方法采用标准长方案,收集2017年至2018年在内蒙古医科大学附属医院生殖医学中心接受体外受精/卵胞质内单精子显微注射(IVF/ICSI)助孕的PCOS患者(PCOS组)和正常排卵女性(对照组)的卵巢颗粒细胞,整合高通量mRNA和miRNA表达谱数据,通过Gene Ontology功能富集和KEGG Pathway分析差异表达基因富集的生物学过程,同时利用miRNA靶向调控基因的关系构建PCOS发病过程中核心调控网络,预测PCOS相关的mRNA与miRNA。结果与对照组相比,PCOS组中66个差异表达miRNA,其中42个表达上调,24个表达下调;mRNA转录谱筛选出416个表达显著差异基因,其中236个基因上调,180个基因下调(P<0.05,|log2FC|≥2)。通过联合分析得到miRNA-26b、miRNA-423-3p、miRNA-219a、miRNA-326、miRNA-3928-3p和miRNA-194-5p关联到107个靶基因,差异有统计学意义(P<0.01)。这些靶基因涉及离子转运、细胞黏附和炎症反应等多方面功能,以及紧密连接丝裂原活化蛋白激酶(MAPK)、Wnt和磷脂酰肌醇-3-激酶/丝氨酸蛋白激酶B(PI3K-Akt)等多个信号通路。结论PCOS患者卵巢颗粒细胞中差异表达的miRNA及其调控的靶基因与其病理生理过程密切相关。

Objective To construct the expression profiles of microRNA(miRNA)and mRNA in ovarian granulosa cells in patients with polycystic ovary syndrome(PCOS)in search of the molecular markers of PCOS.Methods A standard length scheme was used to collect ovarian granulosa cells of PCOS patients(PCOS group)and normal ovulating women(control group)who received in vitro fertilization/intracytoplasmic sperm injection(IVF/ICSI)in the Reproductive Medicine Center of the Affiliated Hospital of Inner Mongolia Medical University from 2017 to 2018,integrate high-throughput mRNA and miRNA expression profiles,and the biological processes involved in differential expression genes were analyzed through Gene Ontogy's functional enrichment and KEGG Pathway.At the same time,the core regulatory network during the pathogenesis of PCOS was constructed by using the relationship between miRNA target regulatory genes,and PCOS-related mRNAs and miRNAs were predicted.Results Compared with control group,66 miRNAs with differential expression were screened in POCS group,among which 42 miRNAs were up-regulated and 24 miRNAs were down-regulated.Totally 416 differentially expressed genes were screened by transcription profiling,among which 236 genes were up-regulated and 180 genes were down-regulated(P<0.05,|log2FC|≥2).Highly significantly differentially expressed miRNAs(miRNA-26b,miRNA-423-3p,miRNA-219a,miRNA-326,miRNA-3928-3p and miRNA-194-5p)were associated with 107 differentially targeted genes by joint analysis,with significant differences(P<0.01).These target genes involve many functions such as ion transport,cell adhesion and inflammatory reactions,and multiple signal pathways such as mitogen-activated protein kinase(MAPK),Wnt and phosphateidy-linositol-3-kinase/serine-threonine kinase(PI3K-Akt).Conclusion The expression of miRNA and its regulatory target genes in ovarian granulosa cells in patients with PCOS are closely related to their pathophysiological processes.