Chen Liping, Xiang Tongwei, Xing Jing, Lu Xinan, Wei Shan, Wang Huaying, Li Jipeng, Yu Wanjun
Department of Central Laboratory, The Affiliated People's Hospital of Ningbo University, Ningbo, People's Republic of China.
Department of Respiratory and Critical Care Medicine, The Affiliated People's Hospital of Ningbo University, Ningbo, People's Republic of China.
Int J Gen Med. 2024 Sep 28;17:4419-4433. doi: 10.2147/IJGM.S473202. eCollection 2024.
It has been increasingly recognized that circular RNAs (circRNAs) act as a pivotal factor in the onset and progression of human malignancies. Yet, the specific activities and mechanistic roles of these RNAs in the context of lung adenocarcinoma (LUAD) are not fully understood.
Microarray analysis identified a novel LUAD-associated circular RNA, termed hsa_circ_0006357 (also referred to as circEZH2). Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was utilized for the analysis of circEZH2 expression in tissues and cell lines. The characteristics of circEZH2 were verified by RNase R treatment and fluorescence in situ hybridization (FISH) assays. The functions of circEZH2 were detected by Cell Counting Kit-8 (CCK-8), colony formation, wound healing, and Transwell assays. The molecular mechanism of circEZH2 was clarified through bioinformatics analysis as well as RNA pulldown, dual-luciferase reporter, RT-qPCR, and immunoblotting assays. The role of circEZH2 in vivo was investigated using a xenograft model.
This investigation revealed that circEZH2 expression was elevated in LUAD cell lines and tumor samples. This elevation was associated with enhanced cell proliferation, migratory capacity, epithelial-mesenchymal transition (EMT), and invasion in vitro. Conversely, silencing of circEZH2 in vivo resulted in a notable decrease in LUAD tumorigenesis, whereas its overexpression led to the opposite effects. Mechanistically, circEZH2 appeared to act as a sponge for miR-495-3p, facilitating the upregulation of tumor protein D52 (TPD52) and triggering the nuclear factor kappa B (NF-κB) signaling pathway, thus contributing to the progression of LUAD.
These findings indicate that circEZH2 may function as a competitive endogenous RNA (ceRNA), driving the progression of LUAD by manipulating the miR-495-3p/TPD52 axis and activating the NF-κB pathway.
环状RNA（circRNAs）在人类恶性肿瘤的发生和发展中作为关键因素已得到越来越多的认识。然而，这些RNA在肺腺癌（LUAD）中的具体活性和机制作用尚未完全明确。
通过微阵列分析鉴定出一种新的与LUAD相关的环状RNA，命名为hsa_circ_0006357（也称为circEZH2）。采用逆转录定量聚合酶链反应（RT-qPCR）分析circEZH2在组织和细胞系中的表达。通过核糖核酸酶R处理和荧光原位杂交（FISH）试验验证circEZH2的特征。采用细胞计数试剂盒-8（CCK-8）、集落形成、伤口愈合和Transwell试验检测circEZH2的功能。通过生物信息学分析以及RNA下拉、双荧光素酶报告基因、RT-qPCR和免疫印迹试验阐明circEZH2的分子机制。使用异种移植模型研究circEZH2在体内的作用。
本研究表明，circEZH2在LUAD细胞系和肿瘤样本中的表达升高。这种升高与体外细胞增殖、迁移能力、上皮-间质转化（EMT）和侵袭增强有关。相反，在体内沉默circEZH2导致LUAD肿瘤发生显著减少，而其过表达则产生相反的效果。机制上，circEZH2似乎作为miR-495-3p的海绵，促进肿瘤蛋白D52（TPD52）的上调并触发核因子κB（NF-κB）信号通路，从而促进LUAD的进展。
这些发现表明，circEZH2可能作为一种竞争性内源性RNA（ceRNA），通过操纵miR-495-3p/TPD52轴并激活NF-κB途径来驱动LUAD的进展。
