Department of Neurosurgery, The People's Hospital of Xinjiang Uygur Autonomous Region, #91 Tianchi Road, Tianshan District, Urumqi, 830001, Xinjiang, China.
Department of Rehabilitative Medicine, The People's Hospital of Xinjiang Uygur Autonomous Region, #91 Tianchi Road, Tianshan District, Urumqi, 830001, Xinjiang, China.
Hum Cell. 2024 Oct 26;38(1):10. doi: 10.1007/s13577-024-01129-z.
Glioblastoma (GBM) is the most aggressive form of glioma, characterized by high mortality and poor prognosis. Dysregulation of microRNAs (miRNAs) plays a critical role in the progression and metastasis of GBM. This study aimed to investigate the role and molecular mechanism of miR-124-3p in GBM. Levels of miR-124-3p, EPHA2, and ALKBH5 were measured using quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation, migration, invasion, and stemness were assessed using the Cell Counting Kit-8 (CCK-8), colony formation, Transwell, and sphere formation assays, respectively. Bioinformatics prediction, dual-luciferase reporter assays, and RNA pull-down experiments were employed to validate the target of miR-124-3p. RNA binding protein immunoprecipitation (RIP) and methylated RNA immunoprecipitation (Me-RIP) were utilized to evaluate the regulation of miR-124-3p maturation by ALKBH5. The results indicated that overexpression of miR-124-3p inhibited the proliferation, migration, invasion, and stemness of GBM cells. EPHA2 was identified as a direct downstream target of miR-124-3p, and its overexpression reversed the inhibitory effects of miR-124-3p on cellular functions. Furthermore, miR-124-3p targeted EPHA2 to inactivate the Wnt/β-catenin pathway. Additionally, ALKBH5 negatively regulated miR-124-3p by impeding its processing. In conclusion, knockdown of ALKBH5 promoted the processing of pri-miR-124-3p, increasing mature miR-124-3p levels, which inhibited the malignant behaviors of GBM cells by targeting EPHA2. These findings highlight the importance of the ALKBH5/miR-124-3p/EPHA2 axis in GBM.
胶质母细胞瘤（GBM）是最具侵袭性的神经胶质瘤，具有高死亡率和预后不良的特点。miRNAs（miRNA）的失调在 GBM 的进展和转移中起着关键作用。本研究旨在探讨 miR-124-3p 在 GBM 中的作用和分子机制。采用实时定量聚合酶链反应（qRT-PCR）检测 miR-124-3p、EPH 受体 A2（EPHA2）和 AlkB 家族成员 5（ALKBH5）的水平。使用细胞计数试剂盒-8（CCK-8）、集落形成、Transwell 和球体形成实验分别评估细胞增殖、迁移、侵袭和干性。利用生物信息学预测、双荧光素酶报告基因检测和 RNA 下拉实验验证 miR-124-3p 的靶基因。采用 RNA 结合蛋白免疫沉淀（RIP）和甲基化 RNA 免疫沉淀（Me-RIP）实验评估 ALKBH5 对 miR-124-3p 成熟的调控作用。结果表明，过表达 miR-124-3p 抑制 GBM 细胞的增殖、迁移、侵袭和干性。EPH 受体 A2（EPHA2）被鉴定为 miR-124-3p 的直接下游靶基因，其过表达逆转了 miR-124-3p 对细胞功能的抑制作用。此外，miR-124-3p 通过靶向 EPHA2 来抑制 Wnt/β-catenin 通路。此外，ALKBH5 通过阻碍其加工来负调控 miR-124-3p。综上所述，敲低 ALKBH5 可促进 pri-miR-124-3p 的加工，增加成熟 miR-124-3p 的水平，通过靶向 EPHA2 抑制 GBM 细胞的恶性行为。这些发现强调了 ALKBH5/miR-124-3p/EPHA2 轴在 GBM 中的重要性。
