Zeng Huan, Huang Mengyi, Gong Xin
Department of Neurosurgery, Hunan Provincial People's Hospital, the First Affiliated Hospital of Hunan Normal University, Changsha, P.R. China.
Neurol Res. 2022 Dec;44(12):1094-1103. doi: 10.1080/01616412.2022.2112374. Epub 2022 Aug 18.
This research was performed to dissect the influence of microRNA (miR)-124-3p on the apoptosis and autophagy of glioma cells and clarify its specific mechanism.
RT-PCR and western blot were utilized to determine miR-124-3p and CREBRF expression in U251 and T98 cells. After loss- and gain-of-function assays in U251 and T98 cells, glioma cell proliferation, autophagy, and apoptosis were measured by MTT assay, western blot, and flow cytometry, respectively. The relationship between miR-124-3p and CREBRF was examined by dual-luciferase reporter assay. The levels of AKT pathway-related proteins were detected by western blot.
MiR-124-3p was lowly expressed and CREBRF was highly expressed in U251 and T98 cells. Overexpression of miR-124-3p or knockdown of CREBRF enhanced apoptosis and autophagy and diminished proliferation of glioma cells. MiR-124-3p negatively targeted CREBRF. MiR-124-3p up-regulation repressed proliferation and facilitated apoptosis and autophagy of glioma cells by diminishing CREBRF expression and blocking the AKT pathway.
MiR-124-3p accelerates apoptosis and autophagy of glioma cells via CREBRF.
本研究旨在剖析微小RNA(miR)-124-3p对胶质瘤细胞凋亡和自噬的影响,并阐明其具体机制。
采用逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法检测U251和T98细胞中miR-124-3p和CREBRF的表达。在U251和T98细胞中进行功能缺失和功能获得实验后,分别通过MTT法、蛋白质免疫印迹法和流式细胞术检测胶质瘤细胞的增殖、自噬和凋亡。采用双荧光素酶报告基因检测法检测miR-124-3p与CREBRF之间的关系。通过蛋白质免疫印迹法检测AKT通路相关蛋白的水平。
miR-124-3p在U251和T98细胞中低表达,CREBRF高表达。miR-124-3p过表达或CREBRF敲低可增强胶质瘤细胞的凋亡和自噬,并减少其增殖。miR-124-3p负向靶向CREBRF。miR-124-3p上调通过降低CREBRF表达和阻断AKT通路抑制胶质瘤细胞增殖,促进其凋亡和自噬。
miR-124-3p通过CREBRF促进胶质瘤细胞的凋亡和自噬。