apicidin对脑胶质瘤U87细胞的影响及对OCT-4基因表达的调控  

Effect of apicidin on glioblastoma U87 cells and its regulation of OCT-4 gene expression

作　　者：李仲颖 王卫红[1] 齐皓 吴炳山[1] 高鹏[1] 汪惊涛 程宏伟[1] Li Zhongying;Wang Weihong;Qi Hao;Wu Bingshan;Gao Peng;Wang Jingtao;Cheng Hongwei(Department of Neurosurgery,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)

机构地区：[1]安徽医科大学第一附属医院神经外科,合肥230022

出　　处：《肿瘤研究与临床》2019年第12期805-808,共4页Cancer Research and Clinic

摘　　要：目的探讨组蛋白去乙酰化酶抑制剂apicidin对脑胶质瘤U87细胞的影响及对OCT-4基因表达的调控作用。方法用不同浓度apicidin处理胶质瘤U87细胞,以二甲基亚砜代替apicidin作为阴性对照。应用四甲基偶氮唑盐(MTT)法检测apicidin处理后U87细胞增殖能力,于共聚焦荧光显微镜下观察细胞凋亡情况,应用流式细胞仪检测细胞周期变化,应用反转录聚合酶链反应和蛋白质印迹法分别检测U87细胞OCT-4的mRNA和蛋白相对于GAPDH的表达量。结果MTT结果显示,apicidin呈时间-剂量依赖性抑制胶质瘤U87细胞增殖,48 h半数抑制浓度为(1.74±0.13)μmol/L。阴性对照及0.2、0.5、1.0μmol/L apicidin作用48 h后U87细胞中S期细胞比例分别为(32.68±0.49)%、(33.73±0.76)%、(42.92±0.56)%、(56.95±0.53)%(P<0.05),而G1、G2期细胞比例减少。共聚焦荧光显微镜下观察,1.0μmol/L apicidin处理48 h后U87细胞出现核固缩等凋亡样改变。与阴性对照组相比,1.0μmol/L apicidin作用48 h后,U87细胞OCT-4 mRNA和蛋白的相对表达量均降低(mRNA:72.44±0.00比56.66±0.23,蛋白:86.59±0.19比56.04±0.15,均P<0.01)。结论apicidin可以抑制脑胶质瘤U87细胞生长,诱导细胞周期阻滞和细胞凋亡,其机制可能与apicidin抑制OCT-4基因的表达有关。Objective To investigate the effect of histone deacetylase inhibitor apicidin on the glioblastoma U87 cells and its regulation of OCT-4 gene expression.Methods Glioblastoma U87 cells were treated with different concentrations of apicidin,and dimethyl sulfoxide instead of apicidin was negative control.Methyl thiazolyl tetrazolium(MTT)assay was used to detect the proliferative ability of U87 cells treated by apicidin.The cell apoptosis was observed under the fluorescence microscope,and the cell cycle was detected by using flow cytometry.Reverse transcription-polymerase chain reaction and Western blot was used to detect the expression of mRNA and protein of U87 cells,respectively relative to the expression of GAPDH.Results MTT assay results showed that apicidin inhibited U87 cells proliferation in a dose-dependent and time-dependent manner,and half of the inhibitory concentration of cell proliferation at 48 h was(1.74±0.13)μmol/L.The cell proportion of U87 cells in S-phase of the negative control,0.2,0.5,and 1.0μmol/L apicidin was(32.68±0.49)%,(33.73±0.76)%,(42.92±0.56)%,and(56.95±0.53)%,respectively after 48 h apicidin administration(P<0.05),while the proportion of G1 and G2 phase cells was decreased.The karyopyknosis and other apoptotic changes were detected in U87 cells after 48 h treatment of 1.0μmol/L apicidin under the confocal fluorescence microscope.Western blot and RT-PCR showed that the mRNA and protein relative levels of U87 cells OCT-4 were reduced after 1.0μmol/L apicidin treatment for 48 h compared with the negative control group(mRNA:72.44±0.00 vs.56.66±0.23;protein:86.59±0.19 vs.56.04±0.15,both P<0.01).Conclusions Apicidin can inhibit the growth of glioblastoma U87 cells,induce cell cycle arrest and apoptosis.Its mechanism may be related to the expression of OCT-4 inhibited by apicidin.

关 键 词：神经胶质瘤 组蛋白脱乙酰基酶类 OCT-4 表观基因组 

分 类 号：R739.4[医药卫生—肿瘤]