Department of Radiotherapy, the Second Affiliated Hospital of Zhengzhou University, Zhengzhou, 450000, Henan, China.
Department of Radiotherapy, the First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Erqi District, Zhengzhou, 450000, Henan, China.
Neurotox Res. 2022 Oct;40(5):1292-1303. doi: 10.1007/s12640-022-00548-w. Epub 2022 Jul 18.
Circular RNA (circRNA) has been shown to be involved in the regulation of human disease progression. Our study aims to reveal the role of circ_0060055 in the progression of glioblastoma (GBM) and its potential molecular mechanism. The expression of circ_0060055, microRNA (miR)-197-3p, and apoptosis inhibitor 5 (API5) was determined by quantitative real-time PCR. GBM cell proliferation, apoptosis, and invasion were assessed using cell counting kit 8 assay, colony formation assay, EdU assay, flow cytometry, and transwell assay. Besides, the radiosensitivity of cells also was assessed using colony formation assay. The interaction between miR-197-3p and circ_0060055 or API5 was analyzed by dual-luciferase reporter assay and RNA pull-down assay. Animal experiments were conducted to measure the effect of circ_0060055 on GBM tumor growth and radiosensitivity in vivo. Circ_0060055 was overexpressed in GBM tumor tissues and cells, and its silencing suppressed GBM cell proliferation and invasion, while promoted apoptosis and radiosensitivity. In terms of mechanism, circ_0060055 could interact with miR-197-3p, and miR-197-3p could target API5. API5 expression also could be positively regulated by circ_0060055. Function experiments suggested that miR-197-3p inhibitor abolished the effect of circ_0060055 knockdown on GBM cell growth, invasion, and radiosensitivity. MiR-197-3p repressed GBM cell progression and improved radiosensitivity, and this effect was eliminated by API5 upregulation. In vivo experiments confirmed that circ_0060055 knockdown reduced GBM tumor growth and enhanced the radiosensitivity of tumors. This study revealed that circ_0060055 contributed to GBM progression and radioresistance through miR-197-3p/API5 pathway, providing a potential target for GBM treatment.
环状 RNA（circRNA）已被证明参与了人类疾病进展的调控。我们的研究旨在揭示 circ_0060055 在神经胶质瘤（GBM）进展中的作用及其潜在的分子机制。通过实时定量 PCR 测定 circ_0060055、微小 RNA（miR）-197-3p 和凋亡抑制剂 5（API5）的表达。通过细胞计数试剂盒 8 测定、集落形成测定、EdU 测定、流式细胞术和 Transwell 测定评估 GBM 细胞增殖、凋亡和侵袭。此外，还通过集落形成测定评估细胞的放射敏感性。通过双荧光素酶报告基因测定和 RNA 下拉测定分析 miR-197-3p 与 circ_0060055 或 API5 的相互作用。进行动物实验以测量 circ_0060055 对体内 GBM 肿瘤生长和放射敏感性的影响。circ_0060055 在 GBM 肿瘤组织和细胞中过表达，其沉默抑制了 GBM 细胞的增殖和侵袭，同时促进了凋亡和放射敏感性。就机制而言，circ_0060055 可以与 miR-197-3p 相互作用，miR-197-3p 可以靶向 API5。API5 的表达也可以被 circ_0060055 正向调节。功能实验表明，miR-197-3p 抑制剂消除了 circ_0060055 敲低对 GBM 细胞生长、侵袭和放射敏感性的影响。miR-197-3p 抑制 GBM 细胞进展并提高放射敏感性，而 API5 的上调则消除了这种作用。体内实验证实，circ_0060055 敲低减少了 GBM 肿瘤的生长并增强了肿瘤的放射敏感性。本研究表明，circ_0060055 通过 miR-197-3p/API5 途径促进 GBM 的进展和放射抵抗，为 GBM 的治疗提供了一个潜在的靶点。
