IGF2BP2抑制剂CWI1-2对胶质瘤细胞迁移、侵袭能力及替莫唑胺耐药性的影响  

Effect of IGF2BP2 inhibitor CWI1-2 on glioma migration,invasion and TMZ resistance

作　　者：伊木然·伊拉木 陶震楠 孙毅 杭春华 Yimuran Yilamu;Tao Zhennan;Sun Yi;Hang Chuhua(Medical School of Nanjing University,Nanjing 210008,China;Department of Neurosurgery,Affiliated Drum Tower Hospital,Medical School of Nanjing University,Nanjing 210000,China;Clinical Medical School of Nanjing Medical University,Nanjing 211166,China)

机构地区：[1]南京大学医学院,南京210008 [2]南京大学医学院附属鼓楼医院神经外科,南京210000 [3]南京医科大学临床医学院,南京211166

出　　处：《中华神经医学杂志》2025年第2期131-140,共10页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金(82203876)。

摘　　要：目的探究胰岛素样生长因子2(IGF2)mRNA结合蛋白2(IGF2BP2)抑制剂CWI1-2对SHG-140、LN229胶质瘤细胞迁移、侵袭能力以及替莫唑胺(TMZ)耐药性的影响。方法(1)体外培养SHG-140、LN229胶质瘤细胞株,采用细胞计数试剂盒-8(CCK-8)实验检测0.1、0.2、0.5、1.0、2.0、5.0μmol/L CWI1-2作用24、48 h对2种细胞存活率的影响,筛选CWI1-2的最佳作用浓度和时间。将2种细胞分为对照组、0.5μmol/L CWI1-2组、1.0μmol/L CWI1-2组,分别加入等量培养基、0.5μmol/L CWI1-2、1.0μmol/L CWI1-2作用24 h,采用蛋白质印迹法和免疫荧光染色检测3组细胞IGF2BP2蛋白的表达,采用细胞划痕实验和Transwell实验分别检测3组细胞的迁移能力、侵袭能力。(2)采用CCK-8实验检测100、200、400、600、800、1000μmol/L TMZ作用24、48 h对SHG-140、LN229细胞存活率的影响,筛选TMZ的最佳作用浓度和时间。将2种细胞分为对照组、CWI1-2组、TMZ组、TMZ+CWI1-2组,分别加入等量培养基、0.5μmol/L CWI1-2、200μmol/L TMZ、0.5μmol/L CWI1-2+200μmol/L TMZ作用24 h,采用细胞集落实验检测4组细胞的集落形成率,采用CCK-8实验检测4组细胞的增殖率。结果(1)CCK-8实验检测结果显示,作用24、48 h后0.5、1.0、2.0、5.0μmol/L CWI1-2组SHG-140、LN229细胞存活率较低于0.1、0.2μmol/L CWI1-2组,差异均有统计学意义(P<0.05),且作用24 h时0.5、1.0、2.0、5.0μmol/L CWI1-2组SHG-140、LN229细胞存活率依次降低,任2组之间差异均有统计学意义(P<0.05)。故选择对细胞存活率影响较小的0.5μmol/L、1.0μmol/L CWI1-2作用24 h用于后续实验。蛋白质印迹法检测结果显示,0.5μmol/L CWI1-2组、1.0μmol/L CWI1-2组SHG-140细胞IGF2BP2蛋白的表达(0.91±0.09、0.79±0.10)低于对照组(1.27±0.05),差异有统计学意义(P<0.05);1.0μmol/L CWI1-2组LN229细胞IGF2BP2蛋白的表达(0.33±0.02)低于对照组(0.82±0.09)和0.5μmol/L CWI1-2组(0.82±0.04),差异有统计学意义(P<0.05)。免疫荧光染色检测Objective To explore the effect of insulin-like growth factor 2(IGF2)mRNA binding protein 2(IGF2BP2)inhibitor CWI1-2 on migration and invasion of glioma cells SHG-140 and LN229 as well as their resistance to temozolomide(TMZ).Methods(1)SHG-140 and LN229 glioma cell lines were cultured in vitro;cell counting kit-8(CCK-8)assay was used to detect the effects of 0.1,0.2,0.5,1.0,2,and 5μmol/L CWI1-2 on survival rate of the two cell lines at 24 and 48 hours after treatment,and the best concentration and treatment time of CWI1-2 were screened.SHG-140 and LN229 cells were categorized into control group,0.5μmol/L CWI1-2 group,and 1.0μmol/L CWI1-2 group,respectively,and equal amount of medium,0.5μmol/L CWI1-2 and 1.0μmol/L CWI1-2 were added into the cells for 24 hours;Western blotting and immunofluorescent staining were used to detect the IGF2BP2 protein expression;cell scratch assay and Transwell assay were,respectively,used to detect the cell migration and invasion.(2)CCK-8 assay was employed to detect the effects of 100,200,400,600,800,and 1000μmol/L TMZ on survival rate of SHG-140 and LN229 cells at 24 and 48 hours after treatment,and the best concentration and treatment time of TMZ were screened.SHG-140 and LN229 cells were categorized into control group,CWI1-2 group,TMZ group,and TMZ+CWI1-2 group,respectively,and equal amounts of culture medium,0.5μmol/L CWI1-2,200μmol/L TMZ,and 0.5μmol/L CWI1-2+200μmol/L TMZ were added into the 4 groups for 24 hours,respectively;colony formation assay was used to detect the colony formation rate and CCK-8 assay was utilized to determine the proliferation rate.Results(1)CCK-8 results indicated that,compared with the 0.1 and 0.2μmol/L CWI1-2 groups,SHG-140 and LN229 cells in the 0.5,1.0,2,and 5μmol/L CWI1-2 groups had significantly lower survival rate at 24 and 48 hours after treatment(P<0.05);furthermore,the survival rate in the SHG-140 and LN229 cells of the 0.5,1.0,2,and 5μmol/L CWI1-2 groups decreased successively,with statistical differences(P<0.05);therefore,0.5μmo

关 键 词：神经胶质瘤 胰岛素样生长因子2 mRNA结合蛋白2 CWI1-2 细胞迁移 细胞侵袭 替莫唑胺 

分 类 号：R739.41[医药卫生—肿瘤]