Yang Xueliang, Sun Quan, Song Yongming, Li Wenli
No.1 Department of Thoracic Surgery, Shanxi Provincial Cancer Hospital, Taiyuan, Shanxi, China.
Int J Genomics. 2021 Dec 3;2021:3997045. doi: 10.1155/2021/3997045. eCollection 2021.
Circular RNAs (circRNAs) are reported as competing endogenous RNAs (ceRNAs) and play key roles in non-small-cell lung cancer (NSCLC) progression. Thus, this study was aimed at clarifying underlying molecular mechanisms of circHUWE1 in NSCLC.
The quantitative real-time polymerase chain reaction (RT-qPCR) and western blot analyses were used for examining circHUWE1, microRNA-34a-5p (miR-34a-5p), and tumor necrosis factor alpha-induced protein 8 (TNFAIP8). IC of cisplatin (DDP) in A549/DDP and H1299/DDP cells and cell viability were analyzed by the Cell Counting Kit 8 (CCK-8) assay. Colony forming assay was performed to assess colony forming ability. Cell apoptosis and cell cycle distribution were determined by flow cytometry. Migrated and invaded cell numbers were examined by transwell assay. The association among circHUWE1, miR-34a-5p, and TNFAIP8 was analyzed by dual-luciferase reporter and RNA immunoprecipitation assays. A xenograft experiment was applied to clarify the functional role of circHUWE1 .
circHUWE1 was upregulated in NSCLC tissues and cells, especially in DDP-resistant groups. circHUWE1 downregulation inhibited DDP resistance, proliferation, migration, and invasion while it induced apoptosis and cell cycle arrest of DDP-resistant NSCLC cells, which was overturned by silencing of miR-34a-5p. TNFAIP8 was a functional gene of miR-34a-5p, and the suppressive effects of miR-34a-5p overexpression on DDP-resistant NSCLC progression were dependent on the suppression of TNFAIP8. circHUWE1 inhibition also delayed tumor growth of DDP-resistant NSCLC cells.
circHUWE1 functioned as a promoter in DDP-resistant NSCLC by interaction with miR-34a-5p-TNFAIP8 networks, providing novel insight into DDP-resistant NSCLC diagnosis and treatment.
环状RNA（circRNAs）被报道为竞争性内源性RNA（ceRNAs），并在非小细胞肺癌（NSCLC）进展中发挥关键作用。因此，本研究旨在阐明circHUWE1在NSCLC中的潜在分子机制。
采用定量实时聚合酶链反应（RT-qPCR）和蛋白质免疫印迹分析来检测circHUWE1、微小RNA-34a-5p（miR-34a-5p）和肿瘤坏死因子α诱导蛋白8（TNFAIP8）。通过细胞计数试剂盒8（CCK-8）检测分析顺铂（DDP）在A549/DDP和H1299/DDP细胞中的半数抑制浓度（IC）以及细胞活力。进行集落形成试验以评估集落形成能力。通过流式细胞术确定细胞凋亡和细胞周期分布。采用Transwell试验检测迁移和侵袭的细胞数量。通过双荧光素酶报告基因和RNA免疫沉淀试验分析circHUWE1、miR-34a-5p和TNFAIP8之间的关联。应用异种移植实验来阐明circHUWE1的功能作用。
circHUWE1在NSCLC组织和细胞中上调，尤其是在顺铂耐药组中。circHUWE1的下调抑制了顺铂耐药性、增殖、迁移和侵袭，同时诱导了顺铂耐药NSCLC细胞的凋亡和细胞周期停滞，而miR-34a-5p的沉默则逆转了这一现象。TNFAIP8是miR-34a-5p的功能基因，miR-34a-5p过表达对顺铂耐药NSCLC进展的抑制作用依赖于对TNFAIP8的抑制。circHUWE1的抑制也延缓了顺铂耐药NSCLC细胞的肿瘤生长。
circHUWE1通过与miR-34a-5p-TNFAIP8网络相互作用，在顺铂耐药NSCLC中发挥促进作用，为顺铂耐药NSCLC的诊断和治疗提供了新的见解。
