Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases, Tianjin Neurosurgical Institute, Tianjin Huanhu Hospital, Tianjin, 300350, China.
Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases, Tianjin Neurosurgical Institute, Tianjin Huanhu Hospital, Tianjin, 300350, China; Department of Neurosurgery, Tianjin Huanhu Hospital, Tianjin, 300350, China.
Biochem Biophys Res Commun. 2018 Sep 18;503(4):2436-2442. doi: 10.1016/j.bbrc.2018.06.173. Epub 2018 Jul 4.
MiR-124-3p and EphA2 are aberrantly expressed in glioma tissue specimens. In the present study, we firstly investigated that miR-124-3p inhibits EphA2 expression mediated by binding its 3'-UTR to regulate the progression of human glioma. The U87MG and LN229 cells were transfected with miR-124-3p mimics and/or siRNA-EphA2, and then the role of miR-124-3p and EphA2 in the colony-formation, cell-cycle, migration and invasion of glioma cells in vitro were examined. Proteins involved in the epithelial-mesenchymal transition were examined using western blot. The results showed that miR-124-3p was significantly downregulated in glioma tissues, whereas a marked upregulation of EphA2 expression was found. Colony-formation and flow cytometry assays demonstrated that EphA2 downregulation or miR-124-3p mimics caused growth and cell-cycle inhibition in glioma. Transwell migration and invasion assays demonstrated that EphA2 downregulation or miR-124-3p mimics suppressed the migration and invasion of glioma cells. EphA2 downregulation or miR-124-3p mimics reduced the level of vimentin in U87MG and LN229 cells. In conclusion, miR-124-3p was found to suppress the growth, migration and invasion of glioma cells in vitro via EphA2. Furthermore, we validated miR-124-3p enforced its biological modulation via targeting EphA2 through the rescue experiment. Conclusively, our study proclaimed that miR-124-3p can counteract the malignant phenotypes of glioma cells by the inhibitory effect of the EphA2.
miR-124-3p 和 EphA2 在胶质瘤组织标本中异常表达。在本研究中，我们首先研究了 miR-124-3p 通过结合其 3'-UTR 抑制 EphA2 表达，从而调节人胶质瘤进展的作用。用 miR-124-3p 模拟物和/或 siRNA-EphA2 转染 U87MG 和 LN229 细胞，然后在体外检测 miR-124-3p 和 EphA2 在胶质瘤细胞集落形成、细胞周期、迁移和侵袭中的作用。使用 Western blot 检测上皮-间充质转化相关蛋白。结果显示，miR-124-3p 在胶质瘤组织中显著下调，而 EphA2 表达明显上调。集落形成和流式细胞术检测表明 EphA2 下调或 miR-124-3p 模拟物导致胶质瘤生长和细胞周期抑制。Transwell 迁移和侵袭实验表明 EphA2 下调或 miR-124-3p 模拟物抑制了胶质瘤细胞的迁移和侵袭。EphA2 下调或 miR-124-3p 模拟物降低了 U87MG 和 LN229 细胞中波形蛋白的水平。结论：miR-124-3p 通过 EphA2 抑制作用抑制胶质瘤细胞的生长、迁移和侵袭。此外，通过挽救实验，我们验证了 miR-124-3p 通过靶向 EphA2 发挥其生物学调节作用。综上所述，我们的研究表明 miR-124-3p 通过抑制 EphA2 来拮抗胶质瘤细胞的恶性表型。
