Wu Dahao, Wang Changzhen
Department of Neurosurgery, Shandong Provincial ENT Hospital, Shandong Provincial ENT Hospital Affiliated to Shandong University, Jinan, China.
Front Neurol. 2020 Apr 28;11:297. doi: 10.3389/fneur.2020.00297. eCollection 2020.
Micro-RNA plays a critical role in the pathological process of gliomas. Previous research showed that the level of miR-155 was significantly increased in many cancers, including gliomas. However, the mechanism of glioma is still unknown. To investigate the regulatory function of miR-155 on glioma U87-MG cells and its effects on related signaling pathways. After transfection of miR-155 mimic and inhibitor, the level of miR-155 were applied to detect cell proliferation, apoptosis, senescence index, invasive ability and cell migration at different time points (0, 24, 24 h, respectively) by CCK8 assay, flow cytometry, β-galactosidase (β-gal) staining, transwell and scratch test, respectively. The effect of miR-155 on PI3K/AKT signal pathway was observed at meantime. Compared with the control group, after miR-155 mimic transfection, U87-MG cell viability, cell migration rate and invasiveness were increased, while apoptosis and senescence were significantly decreased, which was the opposite on miR-155 inhibitor transfection. The phosphorylation levels of miR-155, PI3K, AKT, PI3K, and AKT in U87-MG cells intervened with miR-155 mimic also increased significantly, while the levels of PTEN, Caspase-3, Caspase-9 mRNA, and protein declined significantly, with statistically significant difference. Meanwhile, compared with the control group, miR-155 inhibitor group were on the contrary. The study indicated that miR-155 take charge a key function in regulating the proliferation, migration, and invasion of glioma U87-MG cells through PI3K/AKT signaling pathway, and has anti-glioma effects by inhibition of miR-155, which provided ideas for further clinical treatment of glioma patients.
微小RNA在胶质瘤的病理过程中起关键作用。先前的研究表明,miR-155水平在包括胶质瘤在内的多种癌症中显著升高。然而,胶质瘤的发病机制仍不清楚。为了研究miR-155对胶质瘤U87-MG细胞的调控作用及其对相关信号通路的影响。转染miR-155模拟物和抑制剂后,分别通过CCK8法、流式细胞术、β-半乳糖苷酶(β-gal)染色、Transwell实验和划痕实验,在不同时间点(分别为0、24、48小时)检测miR-155水平,以检测细胞增殖、凋亡、衰老指数、侵袭能力和细胞迁移。同时观察miR-155对PI3K/AKT信号通路的影响。与对照组相比,转染miR-155模拟物后,U87-MG细胞活力、细胞迁移率和侵袭性增加,而凋亡和衰老显著降低,miR-155抑制剂转染后则相反。用miR-155模拟物干预的U87-MG细胞中,miR-155、PI3K、AKT的磷酸化水平也显著增加,而PTEN、Caspase-3、Caspase-9 mRNA和蛋白水平显著下降,差异有统计学意义。同时,与对照组相比,miR-155抑制剂组则相反。研究表明,miR-155通过PI3K/AKT信号通路在调节胶质瘤U87-MG细胞的增殖、迁移和侵袭中起关键作用,抑制miR-155具有抗胶质瘤作用,为胶质瘤患者的进一步临床治疗提供了思路。