ATF3、14-3-3ζ和beclin-1在替莫唑胺治疗敏感与耐药的胶质母细胞瘤中的表达及意义  

Expression of ATF3,14-3-3ζ,and beclin-1 in the treatment of sensitive and drug-resistant glioblastomas with temozolomide and its significance

作　　者：李小燕[1] 赵建祥[2] 任保刚[3] 吴雪晶[1] 吴丹 姚梅宏[1] 陈晓雯 杨映红[1] LI Xiaoyan;ZHAO Jianxiang;REN Baogang;WU Xuejing;WU Dan;YAO Meihong;CHEN Xiaowen;YANG Yinghong(Department of Pathology,Fujian Medical University Union Hospital,Fuzhou 350001;Department of Intensive Care Unit,Fujian Provincial Hospital,Fuzhou 350001;Department of Neurosurgery,Fujian Medical University Union Hospital,Fuzhou 350001,China)

机构地区：[1]福建医科大学附属协和医院病理科,福州350001 [2]福建省立医院重症医学三科,福州350001 [3]福建医科大学附属协和医院神经外科,福州350001

出　　处：《临床与病理杂志》2021年第12期2771-2777,共7页Journal of Clinical and Pathological Research

基　　金：福建省科技创新联合资金项目(2017Y9034);福建省卫生健康科技计划青年项目(2016-1-50);福建医科大学启航基金项目(2016QH021)。

摘　　要：目的:探讨在替莫唑胺(temozolomide,TMZ)治疗敏感与耐药的胶质母细胞瘤中转录激活因子3 (activatingt ranscription factor3,ATF3)、14-3-3ζ和beclin-1的表达水平变化及其与TMZ耐药产生的相关意义。方法:采用RT-PCR及免疫组织化学法检测并比较同一患者TMZ治疗前及耐药时胶质母细胞瘤组织内ATF3、14-3-3ζ和beclin-1基因及蛋白质表达水平。取30μg/m LTMZ培养的U87细胞为实验组,普通溶剂培养的U87细胞为对照组,培养72h,MTT及annexin V-别藻蓝素(allophycoc yanin,APC)单染法分别检测并比较肿瘤细胞的生长抑制率、凋亡率,RT-PCR及蛋白质印迹检测并比较细胞内ATF3、14-3-3ζ和beclin-1基因及蛋白质表达水平。结果:TMZ耐药时胶质母细胞瘤组织中ATF3、14-3-3ζ和beclin-1表达水平均高于TMZ治疗前(P<0.05);TMZ敏感的U87细胞培养72 h肿瘤细胞生长抑制率和凋亡率均增加(P<0.05),细胞内ATF3、14-3-3ζ和beclin-1表达水平均低于对照组(P<0.05);三者呈同向表达。结论:在TMZ用药过程中,胶质母细胞瘤内ATF3激活可能上调14-3-3ζ的表达,进而启动并使自噬增强,导致TMZ耐药产生。Objective:To investigate the changes of the expression of activating transcription factor 3(ATF3),14-3-3ζ,and beclin-1 in the treatment of sensitive and drug-resistant glioblastomas with temozolomide(TMZ).And to investigate their correlation with TMZ resistance.Methods:Before TMZ treatment and after TMZ resistance,the gene and protein expression levels of ATF3,14-3-3ζ,and beclin-1 in glioblastoma tissues of the same patient were detected and compared by RT-PCR and immunohistochemistry.U87 cells cultured with 30 μg/m L TMZ were taken as an experimental group,and U87 cells cultured with common solvent were taken as a control group.After 72 h of culture,the growth inhibition rate and apoptotic rate of tumor cells between the two groups were detected and compared by MTT and annexin V-allophycocyanin(APC) monochrome staining,respectively.After 72 h of culture,the gene and protein expression levels of ATF3,14-3-3ζ,and beclin-1 between the two groups were detected and compared by RT-PCR and Western blotting.Results:The expression of ATF3,14-3-3ζ and beclin-1 in glioblastoma tissues with TMZ resistance was higher than that before TMZ treatment(P<0.05).After 72 h of culture,the growth inhibition rate and apoptotic rate of TMZ-sensitive U87 cells were increased(P<0.05);meanwhile,the expression of ATF3,14-3-3ζ and beclin-1 in TMZ-sensitive U87 cells was lower than that in the control group(P<0.05).All three have the same expression trend.Conclusion:During the TMZ treatment process,the activation of ATF3 in glioblastoma may upregulate the expression of 14-3-3ζ which initiates and enhances the autophagy of tumor cells,leading to TMZ resistance in glioblastoma.

关 键 词：胶质母细胞瘤 替莫唑胺 自噬 转录激活因子3 14-3-3ζ 

分 类 号：R739.4[医药卫生—肿瘤]