背景:环状RNA(circularRNAs,circRNAs)参与肿瘤的发生和发展,包括非小细胞肺癌(NSCLC)。目前,circRNAs在NSCLC中的作用和调控机制尚未完全阐明.本研究旨在探讨circRNAhsa_circ_0008037(circ_0008037)在NSCLC中的作用及调控机制。
方法:通过定量实时聚合酶链反应(RT-qPCR)检测circ_0008037在NSCLC组织和细胞中的表达。进行了功能丧失实验,以分析circ_0008037敲低对增殖的影响,迁移,入侵,和NSCLC细胞的Warburg效应。蛋白质印迹用于蛋白质分析。通过生物信息学分析研究了circ_0008037的调控机制,RNA下拉测定,和双荧光素酶报告基因测定。异种移植试验用于验证circ_0008037在NSCLC中的体内致癌作用。
结果:Circ_0008037在NSCLC组织和细胞中上调。Circ_0008037下调体内肿瘤生长并抑制增殖,迁移,入侵,并降低了体外NSCLC细胞的Warburg效应。机械上,circ_0008037通过海绵作用miR-433-3p调节核普遍存在的酪蛋白激酶和细胞周期蛋白依赖性激酶底物1(NUCKS1)的表达。此外,MiR-433-3p抑制剂逆转了circ_0008037沉默对增殖的抑制作用,迁移,入侵,和NSCLC细胞的Warburg效应。此外,NUCKS1升高推翻了miR-433-3p模拟物对增殖的抑制作用,迁移,入侵,和NSCLC细胞的Warburg效应。
结论:Circ_0008037通过吸附miR-433-3p调节NUCKS1表达,加速肿瘤生长并提高Warburg效应,为非小细胞肺癌治疗提供了潜在的靶点。
BACKGROUND: Circular RNAs (circRNAs) participate in the genesis and progression of tumors, including non-small cell lung cancer (NSCLC). At present, the role and regulatory mechanisms of circRNAs in NSCLC have not been fully elucidated. The aim of this study was to explore the role and regulatory mechanism of circRNA hsa_circ_0008037 (circ_0008037) in NSCLC.
METHODS: Expression of circ_0008037 in NSCLC tissues and cells was detected by quantitative real-time polymerase chain reaction (RT-qPCR). Loss-of-function experiments were performed to analyze the influence of circ_0008037 knockdown on proliferation, migration, invasion, and the Warburg effect of NSCLC cells. Western blotting was utilized for protein analysis. The regulatory mechanism of circ_0008037 was surveyed by bioinformatics analysis, RNA pulldown assay, and dual-luciferase reporter assay. Xenograft assay was used to validate the oncogenicity of circ_0008037 in NSCLC in vivo.
RESULTS: Circ_0008037 was upregulated in NSCLC tissues and cells. Circ_0008037 downregulation reduced tumor growth in vivo and repressed proliferation, migration, invasion, and decreased the Warburg effect of NSCLC cells in vitro. Mechanically, circ_0008037 regulated nuclear ubiquitous casein kinase and cyclin-dependent kinase substrate 1 (
NUCKS1) expression via sponging miR-433-3p. Furthermore, MiR-433-3p inhibitor reversed the inhibiting influence of circ_0008037 silencing on proliferation, migration, invasion, and the Warburg effect of NSCLC cells. Also,
NUCKS1 elevation overturned the repressive influence of miR-433-3p mimic on proliferation, migration, invasion, and the Warburg effect of NSCLC cells.
CONCLUSIONS: Circ_0008037 accelerated tumor growth and elevated the Warburg effect via regulating
NUCKS1 expression by adsorbing miR-433-3p, providing an underlying target for NSCLC treatment.