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微小RNA-124-3p通过下调CREBRF促进胶质瘤细胞的凋亡和自噬。

MicroRNA-124-3p promotes apoptosis and autophagy of glioma cells by down-regulating CREBRF.

作者信息

Zeng Huan, Huang Mengyi, Gong Xin

机构信息

Department of Neurosurgery, Hunan Provincial People's Hospital, the First Affiliated Hospital of Hunan Normal University, Changsha, P.R. China.

出版信息

Neurol Res. 2022 Dec;44(12):1094-1103. doi: 10.1080/01616412.2022.2112374. Epub 2022 Aug 18.

Abstract

OBJECTIVE

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.

METHODS

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.

RESULTS

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.

CONCLUSION

MiR-124-3p accelerates apoptosis and autophagy of glioma cells via CREBRF.

摘要

目的

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。

方法

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。

结果

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。

结论

miR-124-3p通过CREBRF促进胶质瘤细胞的凋亡和自噬。

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