Chu T, Yu T, Wang F, An H L, Shi H, Huang K, Liu Y P, Zhai J
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.
Zhonghua Yi Xue Za Zhi. 2024 Aug 20;104(32):3050-3058. doi: 10.3760/cma.j.cn112137-20240622-01385.
To construct a repetitive implantation failure (RIF)-related competitive endogenous RNA (ceRNA) regulatory network and validate with clinical samples. 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). 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 [ (, )] were 26.50 (25.00, 34.00) and 30.50 (25.75, 35.25) years old, respectively (>0.05). Compared with the control group, the mRNA [0.30 (0.15, 0.42) vs 0.99 (0.69, 1.34), =0.001] and protein expression [0.44 (0.16, 1.27) vs 2.39 (1.58, 2.58), <0.001] of VCAM1 in the endometrium of the RIF group were both reduced. 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.
构建复发性植入失败（RIF）相关的竞争性内源性RNA（ceRNA）调控网络并进行临床样本验证。从高通量基因表达综合数据库（GEO）的表达谱数据集中获取RIF相关的长链非编码RNA（lncRNA）、微小RNA（miRNA）和信使RNA（mRNA），构建lncRNA-miRNA-mRNA的ceRNA调控网络。同时，采用加权基因共表达网络分析（WGCNA）探索该网络中的枢纽基因。本回顾性研究收集了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）/卵胞浆内单精子注射（ICSI）辅助妊娠的RIF患者及对照组（辅助受孕后至少有一次妊娠史）。在有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）验证RIF相关枢纽基因的mRNA表达水平，采用蛋白质印迹法和免疫组织化学法验证血管细胞黏附分子-1（VCAM1）的蛋白表达水平。构建了一个由32个lncRNA、31个miRNA和88个mRNA组成的RIF相关ceRNA调控网络，并通过WGCNA鉴定出7个RIF相关枢纽基因。通过将ceRNA网络中的88个mRNA与枢纽基因进行交叉分析，得到两个RIF相关关键基因，即VCAM1和白细胞介素-2受体α（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[（，）]分别为26.50（25.00，34.00）岁和30.50（25.75，35.25）岁（>0.05）。与对照组相比，RIF组子宫内膜中VCAM1的mRNA[0.30（0.15，0.42）对0.99（0.69，1.34），=0.001]和蛋白表达[0.44（0.16，1.27）对2.39（1.58，2.58），<0.001]均降低。本研究采用生物信息学分析方法构建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的表达水平显著降低。
