Hsa_circ_0003528 通过海绵吸附 miR-511-3p 增加癌基因 PDL1 促进非小细胞肺癌细胞恶性转化和免疫逃逸。
Hsa_circ_0003528 promotes cell malignant transformation and immune escape via increasing oncogene PDL1 through sponging miR-511-3p in non-small cell lung cancer.
机构信息
Department of Oncology, Changsha Central Hospital Affiliated to South China University, Changsha, China.
出版信息
Environ Toxicol. 2023 Jun;38(6):1347-1360. doi: 10.1002/tox.23768. Epub 2023 Mar 22.
BACKGROUND
Accumulating evidence suggests that circular RNAs (circRNAs) play important regulatory roles in non-small cell lung cancer (NSCLC). At present, we aimed to explore the regulatory role of has_circ_0003528 (circ_0003528) in NSCLC.
METHODS
Alterations of circ_0003528 expression in NSCLC samples and cell lines were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Impacts of circ_0003528 on NSCLC cell malignant transformation were analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, transwell invasion, and tube formation assays. Epithelial-mesenchymal transition (EMT)-related markers were detected with western blotting. Pro-inflammatory cytokines were detected by Enzyme-linked immunosorbent assay (ELISA). The regulation mechanism of circ_0003528 was verified by dual-luciferase reporter and RNA pull-down assays. The tumorigenesis role of circ_0003528 was verified by animal experiments.
RESULTS
Higher levels of circ_0003528 were obtained in NSCLC samples and cell lines, and patients with high circ_0003528 expression had a worse prognosis. Silence of circ_0003528 decreased xenograft growth in mouse models and induced cell apoptosis and repressed cell viability, proliferation, invasion, EMT, angiogenesis, and immune escape in NSCLC cells in vitro. Mechanistically, circ_0003528 controlled programmed cell death ligand 1 (PDL1) expression through interaction with miR-511-3p. The inhibiting impacts of circ_0003528 knockdown on NSCLC cell malignant transformation and immune escape were weakened after miR-511-3p silencing. Moreover, PDL1 overexpression partially counteracted miR-511-3p upregulation-mediated suppression on NSCLC cell malignant transformation and immune escape.
CONCLUSIONS
Circ_0003528 facilitated NSCLC cell malignant transformation and immune escape through regulation of the miR-511-3p/PDL1 axis, highlighting the oncogenic role of circ_0003528 in NSCLC.
背景
越来越多的证据表明，环状 RNA（circRNA）在非小细胞肺癌（NSCLC）中发挥重要的调控作用。目前，我们旨在探讨 has_circ_0003528（circ_0003528）在 NSCLC 中的调控作用。
方法
采用实时定量聚合酶链反应（RT-qPCR）检测 NSCLC 样本和细胞系中 circ_0003528 的表达变化。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术、Transwell 侵袭和管形成实验分析 circ_0003528 对 NSCLC 细胞恶性转化的影响。采用 Western blot 检测上皮-间充质转化（EMT）相关标志物。采用酶联免疫吸附试验（ELISA）检测促炎细胞因子。通过双荧光素酶报告和 RNA 下拉实验验证 circ_0003528 的调控机制。通过动物实验验证 circ_0003528 的致瘤作用。
结果
在 NSCLC 样本和细胞系中检测到更高水平的 circ_0003528，circ_0003528 高表达的患者预后较差。circ_0003528 沉默可降低裸鼠模型中的异种移植物生长，并诱导细胞凋亡，抑制 NSCLC 细胞的活力、增殖、侵袭、EMT、血管生成和免疫逃逸。机制上，circ_0003528 通过与 miR-511-3p 相互作用来控制程序性细胞死亡配体 1（PDL1）的表达。沉默 miR-511-3p 后，circ_0003528 敲低对 NSCLC 细胞恶性转化和免疫逃逸的抑制作用减弱。此外，PDL1 过表达部分抵消了 miR-511-3p 上调对 NSCLC 细胞恶性转化和免疫逃逸的抑制作用。
结论
circ_0003528 通过调节 miR-511-3p/PDL1 轴促进 NSCLC 细胞恶性转化和免疫逃逸，突出了 circ_0003528 在 NSCLC 中的致癌作用。
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