实用临床医药杂志2024，Vol.28Issue(19)：10-15,21,7.DOI:10.7619/jcmp.20241064
微小RNA-15a-5p通过RUNX1增强胶质瘤细胞对替莫唑胺的敏感性机制研究
Mechanism of microRNA-15a-5p in enhancing the sensitivity of glioma cells to temozolomide by modulating RUNX1
摘要
Abstract
Objective To investigate the effects of microRNA-15a-5p(miR-15a-5p)on the pro-liferation,invasion and migration of and its mechanism of sensitivity of glioma cells to temozolomide(TMZ).Methods Glioma cell lines H4 and SHG-44,normal astrocytes HA1800 and TMZ-resistant H4 cells were selected.H4 cells were transfected with miR-NC or miR-15a-5p mimics and recorded as the miR-NC group and the miR-15a-5p group,respectively.The cells of miR-15a-5p group was further transfected with Vector or OE-RUNX1 plasmids and treated with 400μmoL/L TMZ,and recorded as miR-15a-5p+Vector group,miR-15a-5p+RUNX1 group,TMZ+Vector group and TMZ+RUNX1 group.Additionally,the miR-NC and miR-15a-5p groups were treated with 400 μmoL/L TMZ,and recorded as TMZ+miR-NC group and TMZ+miR-15a-5p group,respectively.Untreated cells served as the Control group.Cell proliferation was assessed using the Methyl Thiazolyl Tetrazolium(MTT)assay.Invasion and migration were evaluated by Transwell assays.RUNX1 mRNA and miR-15a-5p expression levels were determined by quantitative real-time polymerase chain reaction(RT-PCR).RUNX1 protein expression was analyzed by Western blot.The binding sites between miR-15a-5p and RUNX1 were predicted using the TargetScan online database.The targeting relationship between miR-15a-5p and RUNX1 was validated by dual-luciferase reporter assays.Results RUNX1 mRNA and its protein expression levels were significantly higher in H4 and SHG-44 cells compared to HA1800 cells,while miR-15a-5p expression was significantly lower in H4 and SHG-44 cells(P＜0.000 1).Compared with Control group,the expression level of miR-15a-5p in TMZ-resistant H4 cells was significantly lower(t=18.89,P＜0.000 1);compared with Control group,the expression level of RUNX1 mRNA and protein in TMZ-resistant H4 cells was significantly higher(t=34.11,18.07,P＜0.000 1).Upregulation of miR-15a-5p and TMZ treatment both inhibited cell proliferation,invasion and migration,and the combination of miR-15a-5p upregulation and TMZ significantly enhanced these inhibitory effects.MiR-15a-5p negatively regulated RUNX1 expression.Overexpression of RUNX1 reversed the inhibitory effects of miR-15a-5p upregulation and TMZ on glioma cell proliferation,in-vasion,and migration.Conclusion MiR-15a-5p negatively regulates RUNX1,thereby inhibiting the proliferation,invasion and migration of glioma cells and enhancing their sensitivity to TMZ.关键词
微小RNA-15a-5p/RUNX1基因/胶质瘤/替莫唑胺Key words
microRNA-15a-5p/RUNX1 gene/glioma/temozolomide分类
医药卫生引用本文复制引用
居来提·阿扎提,马涛,王洋,张晶晶..微小RNA-15a-5p通过RUNX1增强胶质瘤细胞对替莫唑胺的敏感性机制研究[J].实用临床医药杂志,2024,28(19):10-15,21,7.基金项目
新疆维吾尔自治区自然科学基金资助项目(2022DO1C503) （2022DO1C503）
