Abstract：
Epithelial mesenchymal transition (EMT) occurring in retinal pigment epithelial cells (RPE) is a crucial mechanism that contributes to the development of age-related macular degeneration (AMD), a pivotal factor leading to permanent vision impairment. Long non-coding RNAs (lncRNAs) have emerged as critical regulators orchestrating EMT in RPE cells. In this study, we explored the function of the lncRNA CYTOR (cytoskeleton regulator RNA) in EMT of RPE cells and its underlying mechanisms. Through weighted correlation network analysis, we identified CYTOR as an EMT-related lncRNA associated with AMD. Experimental validation revealed that CYTOR orchestrates TGF-β1-induced EMT, as well as proliferation and migration of ARPE-19 cells. Further investigation demonstrated the involvement of CYTOR in regulating the WNT5A/NFAT1 pathway and NFAT1 intranuclear translocation in the ARPE-19 cell EMT model. Mechanistically, CHIP, EMSA and dual luciferase reporter assays confirmed NFAT1\'s direct binding to CYTOR\'s promoter, promoting transcription. Reciprocally, CYTOR overexpression promoted NFAT1 expression, while NFAT1 overexpression increased CYTOR transcription. These findings highlight a mutual promotion between CYTOR and NFAT1, forming a positive feedback loop that triggers the EMT phenotype in ARPE-19 cells. These discoveries provide valuable insights into the molecular mechanisms of EMT and its association with AMD, offering potential avenues for targeted therapies in EMT-related conditions, including AMD.
摘要：
视网膜色素上皮细胞（RPE）中发生的上皮间质转化（EMT）是导致年龄相关性黄斑变性（AMD）发展的重要机制。导致永久性视力损害的关键因素。长链非编码RNA（lncRNA）已成为RPE细胞中协调EMT的关键调节因子。在这项研究中,我们探讨了lncRNACYTOR（细胞骨架调节RNA）在RPE细胞EMT中的功能及其潜在机制。通过加权相关网络分析，我们将CYTOR鉴定为与AMD相关的EMT相关lncRNA。实验验证表明，CYTOR协调TGF-β1诱导的EMT，以及ARPE-19细胞的增殖和迁移。进一步的研究表明CYTOR参与调节ARPE-19细胞EMT模型中的WNT5A/NFAT1途径和NFAT1核内易位。机械上,CHIP,EMSA和双荧光素酶报告基因测定证实了NFAT1与CYTOR启动子的直接结合，促进转录。相互,CYTOR过表达促进NFAT1表达，而NFAT1过表达增加CYTOR转录。这些发现强调了CYTOR和NFAT1之间的相互促进，形成了一个正反馈回路，触发ARPE-19细胞中的EMT表型。这些发现为EMT的分子机制及其与AMD的关联提供了有价值的见解，为EMT相关疾病的靶向治疗提供潜在途径，包括AMD。
