多囊卵巢综合征伴胰岛素抵抗患者血清miRNA差异表达及二甲双胍干预作用

目的：二甲双胍对多囊卵巢综合征伴胰岛素抵抗（PCOS-IR）患者血清基因表达谱的影响。方法：选择PCOS-IR患者6例为治疗组,健康育龄期女性6例为健康对照组,采用基因芯片和RT-PCR的方法研究临床诊断为PCOS-IR患者的血清miRNA表达变化及二甲双胍干预后的血清miRNA变化。对筛选并验证后具有差异表达的miRNA先用生物信息学方法,再用Gene Ontology和KEGG通路的数据库进行功能和通路分析。结果：系统聚类结果显示治疗组治疗前与健康对照组和治疗后均存在差异。通过RT-PCR验证,治疗组治疗前与健康对照组比较,得到10个差异表达变化的miRNA（P＜0.05）;治疗组治疗前与同组治疗后比较,得到8个差异表达变化的miRNA（P＜0.05）。GO分析显示：差异表达基因与转录调控、信号转导、跨膜转运、细胞增殖、细胞凋亡、细胞代谢、细胞黏附、蛋白磷酸化、蛋白质运输、多细胞生物发育、凝血功能、神经传导等有关。KEGG分析显示,差异表达miRNA参与显著表达的通路有：EGFR酪氨酸激酶抑制剂抗性、Rap1信号通路、wnt信号通路、前列腺癌通路、mTOR信号通路、cAMP信号通路、磷脂酰肌醇信号系统、MAPK信号通路。共得到878个与PCOS-IR及其二甲双胍治疗更相关的靶基因。结论：部分miRNA参与PCOS-IR的发病和二甲双胍治疗过程。

Study on miRNA expression and target genes in serum of patients with polycystic ovary syndrome and insulin resistance and the intervention of metformin

Objective: To study the effect of metformin on the serum gene expression profiles in polycystic ovary syndrome (PCOS) patients with insulin resistance (IR). Methods: Six PCOS-IR patients were selected for the metformin treatment group and six healthy women of childbearing age as the control group. Gene chip and RT-PCR were used to study the changes of serum miRNA expression in clinically diagnosed PCOS-IR patients and the changes of serum miRNA expression after metformin intervention. For miRNAs with differential expression after screening and validation, bioinformatics method was used before gene ontology and KEGG pathway databases were used for function and pathway analysis. Results: The results of cluster analysis showed that there were differences between the untreated PCOS-IR group and the healthy control group as well as before and after metformin treatment in PCOS-IR patients. RT-PCR validated that there were 10 differentially expressed miRNAs in the untreated PCOS-IR group compared to the healthy control group, and 8 differentially expressed miRNAs before and after metformin treatment in PCOS-IR patients. GO analysis showed that the differentially expressed genes were related to transcription regulation, signal transduction, transmembrane transport, cell proliferation, cell apoptosis, cell metabolism, cell adhesion, protein phosphorylation, protein transport, multicellular biological development, coagulation function, nerve conduction, etc. KEGG analysis showed that the differentially expressed miRNAs were significantly involved in the following pathways: EGFR tyrosine kinase inhibitor resistance, Rap1 signaling pathway, Wnt signaling pathway, prostate cancer pathway, mTOR signaling pathway,cAMP signaling pathway, phosphatidylinositol signaling system, MAPK signaling pathway. With this method to narrow the search scope, 878 genes were obtained and related to insulin resistance and metformin therapy in PCOS patients. Conclusion: Some miRNAs were suggested to be involved in the pathogenesis of insulin resistance in PCOS patients and the treatment process of metformin.