Huang Yan, Zhang Deyu, Zhou Yingfang, Peng Chao
Department of Obstetrics and Gynecology, Peking University First Hospital, Beijing, China.
Biol Reprod. 2025 Jul 21. doi: 10.1093/biolre/ioaf153.
Endometriosis, a chronic and incapacitating gynecological disorder manifesting as severe pelvic pain and infertility, has been increasingly associated with the aberrant regulation of non-coding RNAs (ncRNAs), particularly long non-coding RNAs (lncRNAs) and microRNAs (miRNAs). Despite the growing recognition of their biological significance, the precise molecular mechanisms and functional roles of lncRNAs LINC01140 and LINC01550 in the pathogenesis of endometriosis remain largely unexplored. In this study, we performed an integrative bioinformatics analysis utilizing the Gene Expression Omnibus (GEO) dataset to systematically identify differentially expressed lncRNAs, miRNAs, and messenger RNAs (mRNAs), thereby constructing a comprehensive competing endogenous RNA (ceRNA) regulatory network. Primary endometrial stromal cells were meticulously isolated from ectopic and ovarian endometriotic lesions, and the RNA expression profiles were rigorously validated through RT-PCR. The subcellular localization of lncRNAs was precisely determined using FISH, while the molecular interactions between lncRNAs and miRNAs were elucidated through dual-luciferase reporter assays. To delineate the functional impact, we conducted a series of assays including Western blot analysis to evaluate cellular proliferation, migration, and apoptotic processes. Our findings reveal that the LINC01140/miR-200c-3p and LINC01550/miR-363-3p regulatory networks are markedly upregulated in the cytoplasmic compartment of endometriotic cells, exerting pivotal roles in the progression of the disease. Importantly, the targeted silencing of lncRNAs or the overexpression of miR-363-3p/miR-200c-3p significantly attenuated cellular proliferation and migration, while concurrently inducing apoptosis. These results provide compelling evidence for the critical regulatory mechanisms orchestrated by lncRNAs in endometriosis, thereby establishing a robust theoretical framework for the development of innovative therapeutic interventions targeting ncRNA-mediated pathways.
子宫内膜异位症是一种慢性且使人衰弱的妇科疾病，表现为严重的盆腔疼痛和不孕，它越来越多地与非编码RNA（ncRNA）的异常调控相关，尤其是长链非编码RNA（lncRNA）和微小RNA（miRNA）。尽管人们越来越认识到它们的生物学意义，但lncRNA LINC01140和LINC01550在子宫内膜异位症发病机制中的精确分子机制和功能作用在很大程度上仍未被探索。在本研究中，我们利用基因表达综合数据库（GEO）数据集进行了综合生物信息学分析，以系统地鉴定差异表达的lncRNA、miRNA和信使RNA（mRNA），从而构建一个全面的竞争性内源RNA（ceRNA）调控网络。从异位和卵巢子宫内膜异位病变中精心分离出原代子宫内膜基质细胞，并通过逆转录聚合酶链反应（RT-PCR）严格验证RNA表达谱。使用荧光原位杂交（FISH）精确确定lncRNA的亚细胞定位，同时通过双荧光素酶报告基因检测阐明lncRNA与miRNA之间的分子相互作用。为了描述功能影响，我们进行了一系列检测，包括蛋白质免疫印迹分析，以评估细胞增殖、迁移和凋亡过程。我们的研究结果表明，LINC01140/miR-200c-3p和LINC01550/miR-363-3p调控网络在子宫内膜异位细胞的细胞质区室中显著上调，在疾病进展中发挥关键作用。重要的是，lncRNA的靶向沉默或miR-363-3p/miR-200c-3p的过表达显著减弱细胞增殖和迁移，同时诱导细胞凋亡。这些结果为lncRNA在子宫内膜异位症中所协调的关键调控机制提供了有力证据，从而为开发针对ncRNA介导途径的创新治疗干预措施建立了一个强大的理论框架。
