抑制IGFBP-2基因的表达对U251细胞增殖和凋亡的影响  

Effects of RNA interference silencing IGFBP-2 gene on proliferation and apoptosis of human glioma U251 cells

作　　者：许淑芬[1] 王英凯[2] 马寅芙[1] 刘磊[1] 孙牧男[1] 方艳秋[1] 谭岩[1] 

机构地区：[1]吉林省人民医院中心实验室,吉林长春130021 [2]吉林大学第一医院胃肠内科,吉林长春130021

出　　处：《中国实验诊断学》2014年第10期1572-1575,共4页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省科技厅项目资助课题;编号(201015220)

摘　　要：目的探讨RNA干扰技术沉默胰岛素生长因子结合蛋白2(IGFBP-2)对胶质瘤细胞株U251增殖、凋亡的影响,为胶质瘤的基因治疗提供一个可选择的靶点和方法。方法构建靶向IGFBP-2基因的RNAi表达载体;采用脂质体法将siRNA转染U251细胞,设非特异的siRNA阴性对照组和未转染siRNA的阴性对照组;采用RT-PCR法半定量检测siRNA对IGFBP-2基因表达的抑制作用;用MTT法测定U251细胞增殖变化;流式细胞术检测siRNA诱导细胞凋亡作用。结果构建的RNAi表达载体抑制了IGFBP-2基因的表达(P<0.01);RNA干扰沉默IGFBP-2基因可抑制U251细胞增殖,促进细胞凋亡。结论在细胞水平上,构建的靶向IGFBP-2的siRNA可明显抑制IGFBP-2基因的表达,导致胶质瘤细胞凋亡率明显上升(P<0.01),为进一步利用RNA干扰进行胶质瘤的基因治疗研究奠定基础。Objective To investigate the effect of silencing IGFBP-2 with RNA interference on the proliferation and apoptosis of the glioma U251 cells,and to provide a viable method for the gene therapy of glioma.Methods RNAi ex-pression vector for IFGBP-2 was constructed and transfected to U251 cell with lipofectamin.RT-PCR method was used to detect the variation of IGFBP-2 mRNA level.MTT method was used to measure the variation in the proliferation of U251 cells.Flow cytometer was used to monitor cell apoptosis and changes.Results The construction of RNAi expres-sion vector inhibited the expression of IGFBP-2 mRNA;Silencing IGFBP-2 gene with RNA interference could inhibit the proliferation of U251 cells and induce the apoptosis of U251 cells （P〈0 .0 1 ）.Conclusion RNAi technique can ef-fectively inhibit the expression of IGFBP-2 gene（P〈0.01）,hence inhibit the proliferation of U251 cells and induce the apoptosis of U251 cells.

关 键 词：RNA干扰 胶质瘤 胰岛素生长因子结合蛋白2 增殖 凋亡 

分 类 号：R739.41[医药卫生—肿瘤]