CLIC1对胶质母细胞瘤细胞系凋亡影响的实验研究  

Experimental study on the effect of CLIC1 on apoptosis in glioblastoma cell lines

作　　者：张凯 韩淋 张华军 崔江丽 马勃 王煜 缪星宇[1] ZHANG Kai;HAN Lin;ZHANG Huajun;CUI Jiangli;MA Bo;WANG Yu;MIAO Xingyu(Department of Neurosurgery,Shaanxi Provincial People's Hospital,Xi an 710028,China;不详)

机构地区：[1]陕西省人民医院神经外科,西安710028 [2]空军军医大学唐都医院神经内科

出　　处：《临床神经外科杂志》2025年第1期47-54,共8页Journal of Clinical Neurosurgery

基　　金：陕西省重点研发计划项目(2022SF-166);陕西省自然科学基础研究计划项目(2017JM8103)。

摘　　要：目的研究CLIC1对胶质母细胞瘤(GBM)细胞系U251、T98凋亡的影响。方法根据癌症基因组图谱(TCGA)数据库数据分析CLIC1相关临床表达差异、生存曲线等。设计两条慢病毒序列,稳定敲低U251、T98细胞系中CLIC1表达。平板克隆形成实验、CCK8细胞增殖实验评估对照组、敲低组GBM细胞增殖能力。使用流式检测对照组、敲低组细胞凋亡率。使用免疫荧光、Western Blot检测对照组、敲低组凋亡相关蛋白变化。裸鼠异种移植模型建立,观察CLIC1对裸鼠颅内肿瘤生长的影响、肿瘤大小、凋亡及增殖蛋白染色强度,Western Blot检测鼠脑凋亡蛋白表达量。结果泛癌分析表明,CLIC1在广泛肿瘤中较正常组织高表达,包括胶质瘤,且在胶质瘤中CLIC1表达随级别呈递增趋势,高表达CLIC1的患者整体生存期缩短。敲低CLIC1可抑制GBM细胞克隆形成,降低细胞活力,且能增加GBM细胞系凋亡率,增加凋亡蛋白BAX、Cleaved-Caspase3表达,减少抗凋亡蛋白BCL2表达。体内实验结果表明,CLIC1在维持肿瘤进展中发挥重要作用。结论敲低CLIC1可抑制GBM细胞增殖,促进GBM细胞凋亡和延缓颅内进展。Objective To investigate the effect of CLIC1 on apoptosis in glioblastoma(GBM)cell lines U251 and T98.Methods The clinical expression differences and survival curves related to CLIC1 were analyzed using data from The Cancer Genome Atlas(TCGA)database.Two lentiviral sequences were designed to stably knock down CLIC1 expression in U251 and T98 cell lines.A plate clone formation assay and CCK-8 cell proliferation assay were conducted to evaluate the proliferation ability of GBM cells in control and knockdown groups.The apoptosis rate in these groups was assessed via flow cytometry.Immunofluorescence and Western blotting were employed to detect changes in apoptosis-related protein expression.A nude mouse xenograft model was established to study the impact of CLIC1 on intracranial tumor growth.The tumor size,the intensity of apoptotic and proliferative protein expression were observed.Western blotting was used to detect apoptotic protein expression in the mouse brain.Results Pan-cancer analysis revealed that CLIC1 was significantly overexpressed in various tumors,including gliomas,compared to normal tissues.In gliomas,CLIC1 expression increased with tumor grade,and high CLIC1 expression correlated with shorter overall survival in patients.Knockdown of CLIC1 inhibited GBM cell clone formation,reduced cell viability,and increased the apoptosis rate.Additionally,knockdown enhanced the expression of pro-apoptotic proteins BAX and Cleaved-Caspase3,while decreasing the anti-apoptotic protein BCL2.In vivo experiments demonstrated that CLIC1 played a critical role in sustaining tumor progression.Conclusions Knockdown of CLIC1 inhibits GBM cell proliferation,promotes GBM cell apoptosis,and delays intracranial progression.

关 键 词：胶质母细胞瘤 氯离子胞内通道1 细胞凋亡 

分 类 号：R739.41[医药卫生—肿瘤]