Li Zongyu, Fan Mengdi, Zhou Zhibo, Sang Xianyin
Department of Pulmonary and Critical Care Medicine, Shulan(Hangzhou) Hospital Affiliated to Zhejiang Shuren University Shulan International Medical College, Hangzhou, 310022, Zhejiang, China.
Department of General Practice, Shulan(Hangzhou) Hospital Affiliated to Zhejiang, Shuren University Shulan International Medical College, Hangzhou, 310022, Zhejiang, China.
Mol Biotechnol. 2025 Feb;67(2):484-495. doi: 10.1007/s12033-024-01059-z. Epub 2024 Mar 4.
Circular RNAs (circRNAs) have been identified to be dysregulated in non-small cell lung cancer (NSCLC) and implicated in the progression of this cancer. Here, this work aimed to investigate the role and mechanism of circ_0082374 on NSCLC progression. Levels of circ_0082374, miR-491-5p, GPX4 (glutathione peroxidase 4) and epithelial-mesenchymal transition (EMT)-related proteins were examined by quantitative real-time PCR or western blotting, respectively. Cell proliferation and metastasis were detected using cell counting kit-8, colony formation, EdU, transwell, and Scratch assays. Cell ferroptosis was evaluated by measuring cell survival after the treatment of different ferroptosis inducers or inhibitors, as well as the accumulation of intracellular reactive oxygen species (ROS), ferrous iron (Fe) and malondialdehyde (MDA). The binding between miR-491-5p and circ_0082374 or GPX4 was confirmed using dual-luciferase reporter and RNA pull-down assays. In vivo experiments were conducted using murine xenograft assay and immunohistochemistry. Circ_0082374 was a stable circRNA with high expression in NSCLC tissues and cells. Functionally, circ_0082374 silencing suppressed NSCLC cell proliferation and metastasis. Moreover, its down-regulation enhanced ferroptosis by decreasing iron and lipid peroxidation accumulation. Mechanistically, circ_0082374 could indirectly up-regulate GPX4 expression via miR-491-5p, indicating the circ_0082374/miR-491-5p/GPX4 competitive endogenous RNAs (ceRNA) network. Rescue experiments demonstrated that the miR-491-5p/GPX4 axis mediated the regulatory effects of circ_0082374 exerted on NSCLC cells. Moreover, knockdown of circ_0082374 impeded NSCLC growth and EMT via regulating miR-491-5p and GPX4. Circ_0082374 silencing could suppress NSCLC cell proliferation, metastasis and induce ferroptosis through miR-491-5p/GPX4 axis, suggesting a novel therapeutic approach for NSCLC patients.
环状RNA（circRNAs）已被证实非小细胞肺癌（NSCLC）中表达失调，并与该癌症的进展有关。在此，本研究旨在探讨circ_0082374在NSCLC进展中的作用及机制。分别通过定量实时PCR或蛋白质免疫印迹法检测circ_0082374、miR-491-5p、谷胱甘肽过氧化物酶4（GPX4）以及上皮-间质转化（EMT）相关蛋白的水平。使用细胞计数试剂盒-8、集落形成实验、EdU实验、Transwell实验和划痕实验检测细胞增殖和转移情况。通过测量不同铁死亡诱导剂或抑制剂处理后细胞的存活率，以及细胞内活性氧（ROS）、亚铁离子（Fe）和丙二醛（MDA）的积累来评估细胞铁死亡。使用双荧光素酶报告基因实验和RNA下拉实验证实miR-491-5p与circ_0082374或GPX4之间的结合。采用小鼠异种移植实验和免疫组织化学进行体内实验。Circ_0082374是一种稳定的环状RNA，在NSCLC组织和细胞中高表达。在功能上，circ_0082374沉默可抑制NSCLC细胞增殖和转移。此外，其下调通过减少铁和脂质过氧化积累增强铁死亡。机制上，circ_0082374可通过miR-491-5p间接上调GPX4表达，提示circ_0082374/miR-491-5p/GPX4竞争性内源RNA（ceRNA）网络。挽救实验表明，miR-491-5p/GPX4轴介导了circ_0082374对NSCLC细胞的调控作用。此外，circ_0082374敲低通过调节miR-491-5p和GPX4抑制NSCLC生长和EMT。Circ_0082374沉默可通过miR-491-5p/GPX4轴抑制NSCLC细胞增殖、转移并诱导铁死亡，为NSCLC患者提供了一种新的治疗方法。
