EZH2基因沉默对人脑胶质瘤U251细胞增殖和凋亡影响的研究  

Effects of EZH2 silencing on the apoptosis and proliferation of human brain glioma U251 cells

作　　者：吕佳[1] 孙晓阳[1] 丁涟沭[1] 

机构地区：[1]南京医科大学附属淮安第一医院神经外科,江苏淮安223300

出　　处：《中华肿瘤防治杂志》2012年第7期494-497,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：江苏省医学重点人才基金(RC2007029);江苏省社会发展项目(BS2007037);淮安市科技发展基金(HAS07025);南京医科大学科技发展基金重点项目(06NMUZ047)

摘　　要：目的:研究RNA干扰技术抑制zeste基因增强子人类同源物2(EZH2)的表达对人胶质瘤U251细胞增殖及凋亡的影响。方法:构建靶向EZH2基因的shRNA质粒并转染至U251细胞中,采用RT-PCR和蛋白质印迹法检测EZH2mRNA和蛋白的表达情况,利用四甲基偶氮唑盐(MTT)实验和Annexin V-FITC/PI流式细胞术实验观察转染后U251细胞增殖和凋亡情况。结果:靶向EZH2基因的shRNA质粒成功抑制了U251细胞EZH2基因的表达,mRNA和蛋白表达抑制率分别为56.00%和88.73%;转染shRNA-EZH2后,U251细胞的生长受到明显抑制(P<0.01),在转染96h后,生长抑制率达35.79%;转染48h后,U251细胞早、晚期凋亡率分别为(26.59±0.83)%和(38.63±0.80)%,较阴性质粒组和空白对照组均增加,以晚期明显,差异有统计学意义,P<0.01。结论:EZH2基因的沉默能有效抑制胶质瘤U251细胞的增殖、促进其凋亡,提示EZH2可能成为胶质瘤基因治疗的新靶点。OBJECTIVE： To study the effects of EZH2 gene expression＇s inhibition by RNA interference on the proliferation and apoptosis of human glioma U251 cells. METHODS： The shRNA-expressing plasmid targeting EZH2 gene was constructed and transfected into U251 cells. RT-PCR and Western blot were used to detect the EZH2 gene＇s exprsssion at the level of mRNA and protein; MTT assay was performed for detecting cell proliferation and Annexin V-FITC/PI flow cytometric analysis for cell apoptosis. RESULTS： The shRNA-expressing plasmid targeting EZH2 gene successfully inhibited EZH2 gene＇s expression in U251 cells, and the inhibition rates of the mRNA and protein were 56. 00; and 88.73; seperately. After the transfection of the plasmid, the growth of U251 cells was signifinantly inhibited （P〈0.01）, and the highest growth inhibition rate was 35.79 % in 96 hours. Forty-eight hours after transfection,the early and secondary apoptosis rate of U251 cells were （26.59±0.83）% and （38.63±0.80）% respectively, which were higher than both the blank control group and the negative control group （P〈0.01）. CONCLUSION： The silence of EZH2 gene in glioma U251 cell can inhibit its proliferation, promote its apoptosis, suggesting that EZH2 may become a new target for glioma gene therapy.

关 键 词：RNA干扰 zeste基因增强子人类同源物2 胶质瘤 增殖 凋亡 

分 类 号：R739.4[医药卫生—肿瘤]