摘要
目的构建反复种植失败(RIF)相关竞争性内源RNA(ceRNA)调控网络并进行临床样本验证。方法从高通量基因表达数据库(GEO)得到RIF相关的长链非编码RNA(lncRNA)、微小RNA(miRNA)和信使RNA(mRNA)表达谱数据集,构建lncRNA-miRNA-mRNA的ceRNA调控网络。同时利用加权基因共表达网络分析(WGCNA)探索网络中的枢纽基因。回顾性收集2020—2021年于郑州大学第一附属医院生殖医学中心行体外受精(IVF)/卵胞浆内单精子显微注射(ICSI)助孕的RIF患者和对照组(助孕后至少有1次妊娠史)患者的子宫内膜组织,应用实时荧光定量聚合酶链反应(qRT-PCR)验证RIF相关枢纽基因的mRNA表达水平,并应用Western印迹和免疫组化技术验证血管细胞黏附分子-1(VCAM1)的蛋白表达水平。结果构建了由32个lncRNA、31个miRNA和88个mRNA组成的RIF相关ceRNA调控网络,并利用WGCNA鉴定出7个RIF相关枢纽基因。将ceRNA网络中的88个mRNA与枢纽基因取交集得到2个RIF相关关键基因:VCAM1和白细胞介素2受体α(IL-2RA)。临床验证中,对照组和RIF组的年龄[M(Q_(1),Q_(3))]分别为26.50(25.00,34.00)和30.50(25.75,35.25)岁(P>0.05)。与对照组相比,RIF组子宫内膜中VCAM1的mRNA[M(Q_(1),Q_(3))][0.30(0.15,0.42)比0.99(0.69,1.34),P=0.001]和蛋白表达水平[M(Q_(1),Q_(3))][0.44(0.16,1.27)比2.39(1.58,2.58),P<0.001]均降低。结论本研究成功构建了RIF相关ceRNA调控网络,并通过临床样本证实RIF患者子宫内膜组织中VCAM1的表达水平降低。
Objective To construct a repetitive implantation failure(RIF)-related competitive endogenous RNA(ceRNA)regulatory network and validate with clinical samples.Methods RIF-related long non-coding RNA(lncRNA),microRNA(miRNA)and messenger RNA(mRNA)from the high-throughput gene expression omnibus(GEO)database Expression profile data set were obtained to construct a ceRNA regulatory network of lncRNA-miRNA-mRNA.At the same time,weighted gene co-expression network analysis(WGCNA)was used to explore hub genes in the network.This retrospective study collected RIF patients and controls(at least one pregnancy history after assisted conception)who underwent in vitro fertilization(IVF)/intracytoplasmic sperm injection(ICSI)for assisted pregnancy from 2020 to 2021 at the Reproductive Medicine Center of the First Affiliated Hospital of Zhengzhou University.In the endometrial tissue of patients with 1 pregnancy history,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to verify the mRNA expression levels of RIF-related hub genes,and Western blotting and immunohistochemistry were used to verify protein expression levels of vascular cell adhesion molecule-1(VCAM1).Results A RIF-related ceRNA regulatory network consisting of 32 lncRNAs,31 miRNAs and 88 mRNAs was constructed,and 7 RIF-related hub genes were identified using WGCNA.By intersecting 88 mRNAs and hub genes in the ceRNA network,two RIF-related key genes were obtained,i.e.,VCAM1 and interleukin-2 receptorα(interleukin-2 receptorα,IL-2RA).In clinical verification,the ages of the control group and RIF group[M(Q_(1),Q_(3))]were 26.50(25.00,34.00)and 30.50(25.75,35.25)years old,respectively(P>0.05).Compared with the control group,the mRNA[0.30(0.15,0.42)vs 0.99(0.69,1.34),P=0.001]and protein expression[0.44(0.16,1.27)vs 2.39(1.58,2.58),P<0.001]of VCAM1 in the endometrium of the RIF group were both reduced.Conclusions This study uses bioinformatics analysis methods to construct a RIF-related ceRNA regulatory network,and it is confirmed through clinical samples that the expression level of VCAM1 in the endometrial tissue of RIF patients is significantly reduced.
作者
楚婷
余婷
王芳
安慧玲
史昊
黄凯
刘亚平
翟军
Chu Ting;Yu Ting;Wang Fang;An Huiling;Shi Hao;Huang Kai;Liu Yaping;Zhai Jun(Center for Reproductive Medicine of the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Center for Reproductive Medicine of the Foshan Women and Children Hospital,Foshan 528099,China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2024年第32期3050-3058,共9页
National Medical Journal of China
基金
国家自然科学基金(82071649)
河南省高校重点科研项目(22A320025)
关键词
受精
体外
反复种植失败
非编码RNA
GEO
竞争性内源核糖核酸
枢纽基因
Fertilization in vitro
Repeated implantation failure
Non-coding RNA
Gene expression omnibus
Competitive endogenous RNA
Hub gene
作者简介
楚婷,对本文有同等贡献;余婷,对本文有同等贡献;通信作者:翟军,Email:bestzhai2005@163.com
