[Construction of competitive endogenous RNA regulatory network and clinical verification for repeated implantation failure].

T Chu, T Yu, F Wang, H L An, H Shi, K Huang, Y P Liu, J Zhai
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引用次数: 0

Abstract

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 receptor α (interleukin-2 receptor α, IL-2RA). In clinical verification, the ages of the control group and RIF group [M (Q1, Q3)] 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.

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[竞争性内源性 RNA 调控网络的构建及反复植入失败的临床验证]。
目的构建与重复性植入失败(RIF)相关的竞争性内源性 RNA(ceRNA)调控网络,并用临床样本进行验证。方法:从高通量基因表达总库(GEO)数据库表达谱数据集中获取与RIF相关的长非编码RNA(lncRNA)、microRNA(miRNA)和信使RNA(mRNA),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。这项回顾性研究收集了2020年至2021年在郑州大学第一附属医院生殖医学中心接受体外受精(IVF)/卵胞浆内单精子显微注射(ICSI)辅助妊娠的RIF患者和对照组(至少有一次辅助受孕后妊娠史)。在1次妊娠史患者的子宫内膜组织中,采用实时荧光定量聚合酶链反应(qRT-PCR)检测RIF相关枢纽基因的mRNA表达水平,采用Western印迹和免疫组化检测血管细胞粘附分子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鉴定了7个RIF相关枢纽基因。通过ceRNA网络中88个mRNA与枢纽基因的交叉,得到了两个与RIF相关的关键基因,即VCAM1和白细胞介素-2受体α(interleukin-2 receptor α, IL-2RA)。在临床验证中,对照组和 RIF 组[M(Q1,Q3)]的年龄分别为 26.50(25.00,34.00)岁和 30.50(25.75,35.25)岁(P>0.05)。与对照组相比,mRNA [0.30 (0.15, 0.42) vs 0.99 (0.69, 1.34), P=0.001] 和蛋白质表达 [0.44 (0.16, 1.27) vs 2.39 (1.58, 2.58), PConclusions:本研究利用生物信息学分析方法构建了RIF相关的ceRNA调控网络,并通过临床样本证实VCAM1在RIF患者子宫内膜组织中的表达水平显著降低。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Zhonghua yi xue za zhi
Zhonghua yi xue za zhi Medicine-Medicine (all)
CiteScore
0.80
自引率
0.00%
发文量
400
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