Chang Yajie, Xiang Rui, Guo Qi, Li Jing, Li Xiaolan, Zeng Zhi, Peng Jintao, Liang Xiaoyan
Reproductive Medicine Center, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
GuangDong Engineering Technology Research Center of Fertility Preservation, Guangzhou, China.
Front Genet. 2025 Aug 29;16:1619698. doi: 10.3389/fgene.2025.1619698. eCollection 2025.
Intrauterine adhesion (IUA) is characterized by endometrial fibrosis, posing significant risks to women's reproductive health and fertility. This study aimed to uncover a circRNA-associated ceRNA regulatory network relevant to intrauterine adhesion (IUA), thereby contributing to the understanding of its molecular pathogenesis. The expression data of circRNAs and mRNAs in endometrial tissues of IUA and normal controls were analyzed by RNA sequencing, and microRNAs (miRNAs) expression data was downloaded from GSE165321. Our analysis identified 44 differentially expressed (DE) circRNAs, 41 DEmiRNAs, and 640 DEmRNAs. A comprehensive circRNA-miRNA-mRNA network was constructed using Cytoscape. DEmRNAs were mainly enriched in extracellular matrix structural components, collagen fiber complexes. KEGG pathway analysis further implicated the NF-κB signaling pathway, apoptosis, and Notch signaling in IUA development. A protein-protein interaction network for ceRNA-associated mRNAs was developed through the STRING database, highlighting potential hub genes. To validate these transcriptomic findings, RT-qPCR confirmed significant upregulation of the two leading hub circRNAs, hsa_circ_0000439 and hsa_circ_0000994, in IUA samples compared to normal controls, with results showing consistency with RNA sequencing data (p < 0.05). Functional experiments demonstrated that silencing hsa_circ_0000994 with siRNA significantly decreased the expression levels of fibrosis markers α-SMA and COL1A1 in human endometrial stromal cells treated with TGF-β1. In conclusion, this study presents an in-depth transcriptomic analysis of the aberrantly expressed circRNAs, miRNAs, and mRNAs in endometrial tissues from patients with IUA, culminating in the establishment of a novel circRNA-miRNA-mRNA regulatory network. Hsa_circ_0000994 is likely to play a pivotal role in modulating fibrosis associated with IUA, and represents a promising candidate for targeted therapeutic approaches.
宫腔粘连（IUA）的特征是子宫内膜纤维化，对女性生殖健康和生育能力构成重大风险。本研究旨在揭示与宫腔粘连（IUA）相关的环状RNA（circRNA）相关的竞争性内源RNA（ceRNA）调控网络，从而有助于理解其分子发病机制。通过RNA测序分析了IUA患者和正常对照子宫内膜组织中circRNAs和mRNAs的表达数据，并从GSE165321下载了微小RNA（miRNAs）表达数据。我们的分析鉴定出44个差异表达（DE）的circRNAs、41个DEmiRNAs和640个DEmRNAs。使用Cytoscape构建了一个全面的circRNA-miRNA-mRNA网络。DEmRNAs主要富集在细胞外基质结构成分、胶原纤维复合物中。KEGG通路分析进一步表明NF-κB信号通路、细胞凋亡和Notch信号通路与IUA的发生发展有关。通过STRING数据库构建了ceRNA相关mRNAs的蛋白质-蛋白质相互作用网络，突出了潜在的枢纽基因。为了验证这些转录组学结果，逆转录定量聚合酶链反应（RT-qPCR）证实与正常对照相比，IUA样本中两个主要的枢纽circRNAs，即hsa_circ_0000439和hsa_circ_0000994显著上调，结果与RNA测序数据一致（p < 0.05）。功能实验表明，用小干扰RNA（siRNA）沉默hsa_circ_0000994可显著降低经转化生长因子-β1（TGF-β1）处理的人子宫内膜基质细胞中纤维化标志物α-平滑肌肌动蛋白（α-SMA）和Ⅰ型胶原蛋白α1（COL1A1）的表达水平。总之，本研究对IUA患者子宫内膜组织中异常表达的circRNAs、miRNAs和mRNAs进行了深入的转录组学分析，最终建立了一个新的circRNA-miRNA-mRNA调控网络。Hsa_circ_0000994可能在调节与IUA相关的纤维化中起关键作用，是靶向治疗方法的一个有前景的候选靶点。
