Department of Neurosurgery, Qilu Hospital, Cheeloo College of Medicine and Institute of Brain and Brain-Inspired Science, Shandong University, 107 Wenhua Xi Road, Jinan, 250012, Shandong, China.
Shandong Key Laboratory of Brain Function Remodeling, Qilu Hospital, Jinan, 250012, Shandong, China.
Mol Biol Rep. 2023 Feb;50(2):1333-1347. doi: 10.1007/s11033-022-08063-z. Epub 2022 Dec 2.
The microenvironment of hypoxia is an important factor contributing to the development of glioblastoma (GBM). MicroRNA-588 and its potential target Roundabout-directed receptor 1 (ROBO1) have been reported to promote tumor invasion and proliferation in diseases such as gastric, pancreatic and hepatocellular carcinoma, while their function in GBM and response to hypoxic states remain elusive.
A microarray was leveraged to identify differentially expressed microRNAs in U251 glioma cells cultured under normoxic and hypoxic conditions. The expression of miR-588 was assessed using quantitative real-time PCR (qRT‒PCR). Gain- and loss-of-function studies were used to evaluate the role of miR-588 under hypoxic and normoxic conditions. Cell invasion, migration, proliferation, and vasculogenic mimicry (VM) formation experiments were performed. The relationship between miR-588 and ROBO1 was confirmed using western blot and luciferase reporter assays. Intracranial xenograft tumor mouse models were used to study the function of miR-588 in vivo.
The expression of miR-588 was significantly upregulated in hypoxic glioma cells relative to normoxic glioma cells. miR-588 inhibited the invasive, migratory and VM-forming abilities of glioma cells in vitro and in vivo. Mechanistically, roundabout guidance receptor 1 (ROBO1) is a direct, functionally relevant target of miR-588 in glioma. ROBO1 knockdown suppressed the expression of matrix metallopeptidase 2 (MMP2) and matrix metallopeptidase 9 (MMP9), thereby inhibiting the invasive, migratory and VM-forming abilities of glioma.
MiR-588 regulated the behaviors of hypoxic glioma cells by targeting ROBO1. miR-588 can be used as a prognostic marker for glioma and has potential implications in glioma gene therapy.
缺氧微环境是促进胶质母细胞瘤（GBM）发展的重要因素。miRNA-588 及其潜在靶标 Roundabout 导向受体 1（ROBO1）已被报道在胃癌、胰腺癌和肝癌等疾病中促进肿瘤侵袭和增殖，但其在 GBM 中的功能以及对缺氧状态的反应仍不清楚。
利用微阵列鉴定在常氧和缺氧条件下培养的 U251 神经胶质瘤细胞中差异表达的 microRNAs。使用实时定量 PCR（qRT-PCR）评估 miR-588 的表达。在缺氧和常氧条件下进行 gain- 和 loss-of-function 研究，以评估 miR-588 的作用。进行细胞侵袭、迁移、增殖和血管生成拟态（VM）形成实验。使用 Western blot 和荧光素酶报告基因检测证实 miR-588 与 ROBO1 之间的关系。使用颅内异种移植肿瘤小鼠模型研究 miR-588 在体内的功能。
与常氧神经胶质瘤细胞相比，缺氧神经胶质瘤细胞中 miR-588 的表达明显上调。miR-588 抑制了胶质瘤细胞在体外和体内的侵袭、迁移和 VM 形成能力。机制上，ROBO1 是 miR-588 在胶质瘤中的直接、功能相关靶标。ROBO1 敲低抑制了基质金属蛋白酶 2（MMP2）和基质金属蛋白酶 9（MMP9）的表达，从而抑制了胶质瘤细胞的侵袭、迁移和 VM 形成能力。
miR-588 通过靶向 ROBO1 调节缺氧胶质瘤细胞的行为。miR-588 可作为胶质母细胞瘤的预后标志物，并具有胶质母细胞瘤基因治疗的潜力。
