慢病毒载体介导的E2F-1基因沉默对人胶质瘤U251细胞生物学特征的影响  

Effect of lentiviral vector mediated E2F-1 gene silencing on biological characteristics of human glioma U251 cells

作　　者：郭宝光 李虹[1] 李婷[1] 王军[1] 仇波[1] GUO Bao-guang;LI Hong;LI Ting;WANG Jun;QIU Bo(Department of Neurosurgery,the First Affiliated Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院神经外科,辽宁沈阳110001

出　　处：《解剖科学进展》2018年第6期622-625,共4页Progress of Anatomical Sciences

基　　金：国家自然科学基金青年基金(81000565/H0914);沈阳市科技计划项目(18-013-0-60)

摘　　要：目的探讨慢病毒载体介导的E2F-1基因沉默对人胶质瘤U251细胞生物学特征的影响。方法构建靶向E2F-1的shRNA重组慢病毒载体,将慢病毒载体转染U251细胞,real-time PCR与Western blot在m RNA与蛋白水平鉴定转染结果,MTT方法检测转染后U251细胞活力的变化,流式细胞术检测细胞周期及细胞凋亡的变化,transwell实验检测细胞侵袭能力的变化。结果 real-time PCR方法与Western blot检测结果均表明成功建立了稳定沉默E2F-1基因的U251细胞株。与空白对照组及阴性对照组比较,转染E2F-1-shRNA的U251细胞活力显著降低,细胞周期被阻滞于G0/G1期,细胞凋亡率显著升高,侵袭能力显著降低(P<0.01)。结论转染E2F-1-shRNA能够显著降低U251细胞增殖侵袭能力。Objective To investigate the effect of lentiviral vector mediated E2 F-1 gene silencing on the biological characteristics of human glioma U251 cells. Methods The recombinant lentivirus vector of shRNA targeted E2 F-1 was constructed. The lentivirus vector was transfected into U251 cells, real-time PCR and Western blot were used to identify the transfection results at the level of mRNA and protein. MTT method was used to detect the activity of U251 cells after the transfection. Flow cytometry was used to detect the changes of cell cycle and apoptosis. Transwell assay was used to detect the changes of cell invasiveness. Results U251 cell lines with stable E2 F-1 gene were successfully established. Compared with the blank control group and the negative control group, the activity of U251 cells in the E2 F-1-shRNA group was decreased significantly, the cell cycle was blocked in the G0/G1 stage, the apoptosis rate of U251 cells was increased significantly, and the invasion ability of U251 cells was decreased significantly(P<0.01). Conclusion Transfection of E2 F-1-shRNA could significantly reduce the proliferation and invasion ability of U251 cells.

关 键 词：胶质瘤 慢病毒载体 E2F-1基因 U251细胞 

分 类 号：R739.41[医药卫生—肿瘤]