Department of Otorhinolaryngology, Head and Neck Surgery, Zigong First People's Hospital, No.42, Shangyihao Yi Zhi Road, Ziliujing, Zigong, 643000, Sichuan, China.
Mol Cell Biochem. 2021 Aug;476(8):2951-2961. doi: 10.1007/s11010-021-04129-x. Epub 2021 Mar 23.
Circular RNAs (circRNAs) are implicated in the tumorigenesis of human cancers. However, the effects of circRNAs on laryngeal squamous cell carcinoma (LSCC) are largely unknown. Here, we aimed to explore the roles of circ_0023028 in LSCC development. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to measure circ_0023028, miR-486-3p, and Lin-Isl-Mec (LIM) and SH3 domain protein 1 (LASP1) mRNA. The characteristics of circ_0023028 were determined by RNase R digestion assay and Actinomycin D assay. Cell Counting Kit-8 (CCK-8) assay and colony formation assay were utilized for cell proliferation. Transwell assay was adopted for cell invasion and migration. Flow cytometry analysis was carried out to analyze cell cycle and apoptosis. RNA pull-down assay and dual-luciferase reporter assay were used to explore the association between miR-486-3p and circ_0023028 or LASP1. Western blot assay was adopted to measure LASP1 protein level. Murine xenograft model was executed to investigate the function of circ_0023028 in vivo. High expression of circ_0023028 was observed in LSCC tissues and cells. Circ_0023028 was stable and possessed a loop structure. Circ_0023028 silencing suppressed LSCC cell proliferation, metastasis and cell cycle process and induced apoptosis in vitro and hampered tumor growth in vivo. Further mechanism analysis demonstrated that circ_0023028 could sponge miR-486-3p to regulate LASP1 expression in LSCC cells. The malignant behaviors of LSCC cells mediated by circ_0023028 knockdown were rescued by the inhibition of miR-486-3p. Moreover, miR-486-3p suppressed LSCC cell progression via binding to LASP1. Circ_0023028 knockdown might impede the progression of LSCC by regulating miR-486-3p/LASP1 axis, which could provide a novel insight on the mechanism of LSCC progression.
环状 RNA（circRNAs）参与了人类癌症的肿瘤发生。然而，circRNAs 对喉鳞状细胞癌（LSCC）的影响在很大程度上尚不清楚。在这里，我们旨在探索 circ_0023028 在 LSCC 发展中的作用。采用实时定量聚合酶链反应（qRT-PCR）测定 circ_0023028、miR-486-3p、Lin-Isl-Mec（LIM）和 SH3 结构域蛋白 1（LASP1）mRNA。通过 RNase R 消化试验和 Actinomycin D 试验确定 circ_0023028 的特征。细胞计数试剂盒-8（CCK-8）试验和集落形成试验用于细胞增殖。Transwell 试验用于细胞侵袭和迁移。流式细胞术分析用于细胞周期和凋亡分析。RNA 下拉试验和双荧光素酶报告基因试验用于探讨 miR-486-3p 与 circ_0023028 或 LASP1 的关联。Western blot 试验用于测量 LASP1 蛋白水平。建立了小鼠异种移植模型以在体内研究 circ_0023028 的功能。在 LSCC 组织和细胞中观察到 circ_0023028 的高表达。Circ_0023028 稳定且具有环结构。Circ_0023028 沉默抑制 LSCC 细胞的体外增殖、转移和细胞周期过程并诱导细胞凋亡，并阻碍体内肿瘤生长。进一步的机制分析表明，circ_0023028 可以通过海绵 miR-486-3p 来调节 LSCC 细胞中的 LASP1 表达。circ_0023028 敲低介导的 LSCC 细胞恶性行为可以通过抑制 miR-486-3p 得到挽救。此外，miR-486-3p 通过与 LASP1 结合抑制 LSCC 细胞的进展。Circ_0023028 敲低可能通过调节 miR-486-3p/LASP1 轴来阻碍 LSCC 的进展，这为 LSCC 进展的机制提供了新的见解。
