miR-324-3p/MAGOH分子轴在脑胶质瘤中的功能及作用机制研究

Study of the Function and Mechanism of miR-324-3p/MAGOH Molecular Axis in Brain Glioma

目的探究miR-324-3p/MAGOH分子轴在脑胶质瘤中的功能及作用机制。方法收集2022年6月—2023年6月遵义医科大学附属医院神经外科接受手术治疗的胶质瘤患者(n=50)和非胶质瘤的脑病患者(n=20)样本,体外培养人脑胶质瘤细胞系U87。实时荧光定量核酸扩增检测系统(Real-time Quantitative PCR Detecting System,qPCR)检测脑胶质瘤细胞(U87)和患者临床标本中miR-324-3p和MAGOH的表达。蛋白免疫印迹法(Western Blot,WB)检测MAGOH、基质金属蛋白酶(matrix metalloproteinase,MMP)2和MMP9的表达水平。细胞计数试剂盒(Cell Counting Kit-8,CCK-8)检测胶质瘤细胞的增殖能力。Transwell检测胶质瘤细胞的侵袭能力。荧光素酶报告基因检测miR-324-3p和MAGOH的相互作用。免疫组化检测MAGOH在脑胶质瘤组织中的表达。结果胶质瘤患者癌组织中MAGOH的表达较高,差异有统计学意义(P<0.01);且与miR-324-3p的表达呈现显著负相关(r=0.7383,P=0.0002)。过表达MAGOH能促进胶质瘤细胞的增殖和侵袭(P<0.01),而过表达miR-324-3p能抑制胶质瘤细胞的增殖和侵袭(P<0.01)。miR-324-3p和MAGOH的3’-UTR相互作用,且过表达MAGOH能抑制miR-324-3p对胶质瘤细胞的抑制作用(P<0.01)。结论miR-324-3p通过抑制MAGOH的表达来抑制胶质瘤细胞和增殖和侵袭。

Objective To investigate the function and mechanism of miR-324-3p/MAGOH molecular axis in brain glioma.Methods Samples of glioma patients(n=50)and non-glioma encephalopathy patients(n=20)who received sur‐gical treatment in the Department of Neurosurgery of Affiliated Hospital of Zunyi Medical University from June 2022 to June 2023 were collected,and human glioma cell line U87 was cultured in vitro.the real-time quantitative PCR de‐tecting system(qPCR)detects the expression of miR-324-3p and MAGOH in glioma cells(U87)and clinical speci‐mens.Western blot(WB)was used to detect the expression of MAGOH,matrix metalloproteinase(MMP)2 and MMP9.Cell counting kit-8(CCK-8)is used to detect the proliferation of glioma cells.The invasive ability of glioma cells was measured by Transwell.Luciferase reporter gene to detect the interaction between miR-324-3p and MAGOH.The ex‐pression of MAGOH in brain glioma was detected by immunohistochemistry.Results The expression of MAGOH in cancer tissues of glioma patients was higher,and the difference was statistically significant(P<0.01).There was a sig‐nificant negative correlation with the expression of miR-324-3p(r=0.7383,P<0.001).Overexpression of MAGOH could promote the proliferation and invasion of glioma cells(P<0.01),while overexpression of miR-324-3p could in‐hibit the proliferation and invasion of glioma cells(P<0.01).miR-324-3p interacts with the 3'-UTR of MAGOH,and overexpression of MAGOH can inhibit the inhibitory effect of miR-324-3p on glioma cells(P<0.01).Conclusion miR-324-3p inhibited glioma cells and proliferation and invasion by suppressing the expression of MAGOH.