Department of Pulmonary and Critical Care Medicine, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, China.
Department of Clinical Oncology, Shengjing Hospital of China Medical University, No. 39 Huaxiang Road, 110022, Tiexi District, Shenyang, Liaoning Province, China.
Strahlenther Onkol. 2023 Feb;199(2):169-181. doi: 10.1007/s00066-022-01965-8. Epub 2022 Jun 24.
Circular RNAs (circRNAs) have been reported to be crucial modulatory molecules in the etiology of non-small cell lung cancer (NSCLC). This study aimed to probe the precise role and mechanism of circRNA discs large MAGUK scaffold protein 1 (circDLG1) in the malignant progression of NSCLC.
The abundances of circDLG1, miR-630, and centromere protein F (CENPF) mRNAs were gauged by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was tested in 3‑(4, 5‑dimethylthiazol-2-yl)-2, 5‑diphenyltetrazolium bromide (MTT) assay and 5‑ethynyl-2'-deoxyuridine (EdU)-incorporation assay. Cell apoptosis was analyzed by flow cytometry. Cell migration and invasion were assessed by transwell assay. Western blot was exploited to examine the levels of all proteins. The interaction between miR-630 and circDLG1 or CENPF was verified by dual-luciferase reporter, RNA pull-down, and/or RNA immunoprecipitation assays. Tumor xenograft assay and immunohistochemistry (IHC) were executed for the role of circDLG1 in tumor growth in vivo.
CircDLG1 and CENPF were highly expressed in NSCLC, while miR-630 was downregulated. CircDLG1 silencing repressed proliferation, migration, and invasion, and expedited apoptosis of NSCLC cells in vitro. Mechanistically, circDLG1 deficiency modulated NSCLC cell malignant development through interacting with miR-630. Furthermore, CENPF was targeted by miR-630, and circDLG1 could positively control CENPF expression through acting as an miR-630 sponge. Furthermore, CENPF overexpression reversed the repressive impacts of circDLG1 inhibition in the malignant behaviors of NSCLC cells. Besides, circDLG1 interference hindered tumor growth in vivo.
CircDLG1 knockdown could impede NSCLC advancement through modulating the miR-630/CENPF axis, manifesting as a promising molecular target for NSCLC treatment.
环状 RNA（circRNA）已被报道在非小细胞肺癌（NSCLC）的发病机制中是重要的调节分子。本研究旨在探讨 circRNA discs large MAGUK 支架蛋白 1（circDLG1）在 NSCLC 恶性进展中的精确作用和机制。
采用实时定量聚合酶链反应（qRT-PCR）检测 circDLG1、miR-630 和着丝粒蛋白 F（CENPF）mRNA 的丰度。通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）试验和 5-乙炔基-2'-脱氧尿苷（EdU）掺入试验检测细胞增殖。通过流式细胞术分析细胞凋亡。通过 Transwell 试验评估细胞迁移和侵袭。采用 Western blot 检测所有蛋白的水平。通过双荧光素酶报告、RNA 下拉和/或 RNA 免疫沉淀试验验证 miR-630 与 circDLG1 或 CENPF 的相互作用。进行肿瘤异种移植试验和免疫组织化学（IHC）检测 circDLG1 在体内肿瘤生长中的作用。
circDLG1 和 CENPF 在 NSCLC 中高表达，而 miR-630 下调。circDLG1 沉默抑制 NSCLC 细胞的体外增殖、迁移和侵袭，并促进细胞凋亡。机制上，circDLG1 缺失通过与 miR-630 相互作用调节 NSCLC 细胞的恶性发展。此外，CENPF 是 miR-630 的靶标，circDLG1 可以通过作为 miR-630 的海绵正向控制 CENPF 的表达。此外，CENPF 的过表达逆转了 circDLG1 抑制对 NSCLC 细胞恶性行为的抑制作用。此外，circDLG1 干扰抑制体内肿瘤生长。
circDLG1 敲低可通过调节 miR-630/CENPF 轴抑制 NSCLC 进展，有望成为 NSCLC 治疗的有前途的分子靶点。
