摘要
目的通过构建多囊卵巢综合征(polycystic ovary syndrome,PCOS)患者卵丘颗粒细胞的环状RNA(circular RNA,circRNA)-微小RNA(microRNA,miRNA)-信使RNA(messenger RNA,mRNA)调控网络,并进行临床标本验证,以探寻PCOS发病机制并提供诊疗靶点。方法使用R软件分析来自基因表达图谱(gene expression omnibus,GEO)数据库的数据,得到差异表达的circRNA、miRNA、mRNA;使用CircInteractome、Starbase数据库,预测构建circRNA-miRNA-mRNA调控网络。回顾性研究收集2018年1月至2018年12月期间于郑州大学第二附属医院生殖医学部就诊的PCOS患者(记为PCOS组,n=40)和因男方或者输卵管因素治疗的患者(记为对照组,n=20)的卵丘颗粒细胞,提取RNA后进行实时荧光定量PCR(real time quantitative PCR,RT-qPCR)验证。结果筛选出278个差异表达mRNA、23个差异miRNA和2402个差异circRNA(P<0.05和|log_(2)FC|>0.8);构建256条circRNA-miRNA-mRNA调控网络,包含13个mRNA、2个miRNA和40个circRNA;临床验证后提示,妊娠相关血浆蛋白A(pregnancy-associated plasma protein A,PAPPA)、hsa-miR-127-3p、hsairc086809/hsairc063556及其调控网络与PCOS相关(P=0.004、P=0.002、P=0.014、P=0.003)。结论卵丘颗粒细胞中PAPPA、hsa-miR-127-3p、hsairc086809/hsairc063556的表达水平及其调控网络与PCOS发生相关。
Objective To identify the differentially expressed genes,construct circular RNA-microRNA-messenger RNA(cirRNA-miRNA-mRNA)regulatory network,and detect the differentially expressed genes with the clinical samples from patients with polycystic ovary syndrome(PCOS)for further investigating of the mechanisms of pathogenesis and providing novel biomarkers for PCOS,to further explore the pathogenesis of PCOS and provide therapeutic targets.Methods The software‘R’was used to analyze the data from gene expression omnibus(GEO).The differentially expressed genes(mRNA,miRNA and circRNA)were identified,and the mRNA-miRNA-cirRNA regulatory network was predicted by CircInteractome and Starbase database.The retrospective study was performed based on the PCOS patients in the Department of Reproductive Medicine,the Second Affiliated Hospital of Zhengzhou University during January to December in 2018.The cumulus cells were collected from the PCOS(named PCOS group,n=40)and healthy women(named control group,n=20).Reverse transcription real time quantitative PCR(RT-qPCR)was performed to further detect and verify the differentially expressed genes and the network.Results Analysis from GEO database identified the differentially expressed genes including 278 mRNAs,23 miRNAs and 2402 circRNAs in PCOS group compared with non-PCOS group(P<0.05 and|log_(2)FC|>0.8);256 mRNA-miRNA-circRNA regulatory networks were established with the differentially expressed genes including 13 mRNAs,2 miRNAs and 40 circRNAs from the database analysis.The verification with the clinical samples finally revealed the regulatory networks of mRNA pregnancy-associated plasma protein A(PAPPA)-miRNA(hsa-miR-127-3p)-circRNA(hsa_circ_0086809/hsa_circ_0063556)were associated with PCOS(P=0.004,P=0.002,P=0.014,P=0.003).Conclusion The expressions of mRNA(PAPPA),miRNA(hsa-miR-127-3p)and circRNA(hsa_circ_0086809/hsa_circ_0063556)in the cumulus cells and their regulatory networks were associated with PCOS.
作者
马丽影
张乐
许晓婷
窦倩
范晓明
项云改
李朋粉
谭丽
Ma Liying;Zhang Le;Xu Xiaoting;Dou Qian;Fan Xiaoming;Xiang Yungai;Li Pengfen;Tan Li(Department of Reproductive Medicine,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)
出处
《中华生殖与避孕杂志》
CAS
CSCD
北大核心
2022年第5期484-489,共6页
Chinese Journal of Reproduction and Contraception
基金
河南省医学科技攻关计划项目(LHGJ20200421,LHGJ20190318)。
