miR-4488靶向SCRT1调控胶质母细胞瘤细胞的迁移和侵袭  

MiR-4488 regulates migration and invasion of glioblastoma cells by targeting SCRT1

作　　者：谭筱蓉 徐超 汪攀 吴南 TAN Xiaorong;XU Chao;WANG Pan;WU Nan(Chongqing Medical University,Chongqing 400016,China;Department of Neurosurgery,Chongqing General Hospital,Chongqing 401147,China)

机构地区：[1]重庆医科大学,重庆400016 [2]重庆市人民医院神经外科,重庆401147

出　　处：《重庆医学》2025年第4期806-812,共7页Chongqing Medical Journal

基　　金：重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX0548)。

摘　　要：目的探究微RNA-4488(miR-4488)通过调节scratch家族转录抑制因子1(SCRT1)的表达水平影响胶质母细胞瘤(GBM)增殖、迁移及侵袭能力的分子机制。方法采用实时荧光定量PCR(qPCR)技术检测星形胶质细胞SVG和GBM U87MG中miR-4488及SCRT1的表达水平;通过瞬时转染技术将miR-4488 mimic nc(模拟物对照)、miR-4488 mimic(模拟物组)、miR-4488 inhibitor nc(抑制剂对照组)、miR-4488 inhibitor(抑制剂组)分别导入U87MG细胞并相应分为4组;利用慢病毒转染技术将构建的SCRT1空载质粒和SCRT1过表达质粒转染至U87MG细胞系,分别命名为对照组和过表达组。运用生物信息学方法分析并确认miR-4488与SCRT1的结合位点序列,并通过双荧光素酶报告基因实验验证miR-4488对SCRT1的靶向调控关系。利用EdU实验评估各组细胞增殖能力,Transwell实验分析各组细胞迁移与侵袭能力差异。结果与SVG细胞比较,U87MG细胞中miR-4488表达水平上调(P<0.001),SCRT1表达水平下调(P<0.001);瞬时转染后,模拟物组SCRT1表达水平较模拟物对照组下降,增殖能力未见明显变化(P>0.05),但迁移、侵袭能力增强(P<0.01,P<0.001);与之相反,抑制剂组SCRT1表达水平较抑制剂对照组上升,增殖能力未见明显变化,而迁移、侵袭能力减弱(P<0.01,P<0.001)。双荧光素酶报告基因实验证实U87MG细胞中SCRT1是miR-4488的作用靶点。慢病毒转染后,SCRT1过表达组较对照组的迁移、侵袭能力减弱(P<0.01,P<0.001)。结论miR-4488可特异性调控SCRT1的表达,进而影响GBM的迁移和侵袭特性。Objective To investigate the specific molecular mechanism through which microRNA-4488(miR-4488)regulates the proliferation,migration and invasion capabilities of glioblastoma(GBM)by modulating the expression level of scratch family transcriptional repressor 1(SCRT1).Methods Quantitative real-time PCR(qPCR)was performed to measure the expression levels of miR-4488 and SCRT1 in astrocyte SVG cells and GBM U87MG cells.Transient transfection was used to introduce miR-4488 mimic nc(mimic control group),miR-4488 mimic(mimic group),miR-4488 inhibitor nc(inhibitor control group),and miR-4488 inhibitor(inhibitor group)into U87MG cells,which were then divided into four groups accordingly.Lentiviral transfection was used to establish U87MG cell lines transfected with SCRT1 empty vector(control group)and SCRT1 overexpression plasmid(overexpression group).Bioinformatics analysis was performed to identify and validate the binding site sequence between miR-4488 and SCRT1,and the dual-luciferase reporter gene assay was conducted to verify their targeting relationship.The EdU assay was employed to assess cell proliferation capacity,while the Transwell assay was used to analyze differences in migratory and invasive capacities among groups.Results Compared with SVG cells,miR-4488 expression was upregulated(P<0.001)and SCRT1 expression was downregulated in U87MG cells(P<0.001).After transient transfection with miR-4488 mimic,the expression of SCRT1 in the mimic group decreased compared to the mimic control group,with no significant change in proliferative capacity(P>0.05),but enhanced migration and invasion abilities(P<0.01 and P<0.001,respectively).Conversely,after transfection with miR-4488 inhibitor,the expression of SCRT1 in the inhibitor group increased compared to the inhibitor control group,with no significant change in proliferative capacity(P>0.05),but weakened migration and invasion abilities(P<0.01 and P<0.001,respectively).The dual-luciferase reporter gene assay confirmed that SCRT1 is a target of miR-4488 in U87MG cells

关 键 词：胶质母细胞瘤 微RNA SCRT1 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]