慢病毒介导TIEG1基因乳腺癌靶向载体的构建及活性测定  

Construction of lentiviral vector expressing TIEG1 gene with breast cancer specific promoter and identification of activity

作　　者：蒋磊[1] 林飞燕[1] 王福乐[1] 叶爱芳[1] 吴建波[1] 

机构地区：[1]温州医学院附属第一医院内科实验室,325000

出　　处：《实用医学杂志》2011年第11期1905-1908,共4页The Journal of Practical Medicine

基　　金：温州市科技局科研计划资助项目(编号:Y20100017);浙江省教育厅科研计划资助项目(编号:Y200906317)

摘　　要：目的:构建慢病毒介导的TIEG1基因乳腺癌特异性靶向载体,并鉴定其活性。方法:将乳腺癌特异性survivin启动子片段和TIEG1基因先后克隆进带有绿色荧光蛋白的慢病毒载体中,构建survivin启动子驱动的TIEG1基因慢病毒表达载体,酶切及测序鉴定。包装纯化慢病毒颗粒,感染人脐静脉内皮细胞HUVEC和乳腺癌细胞SK-BR-3,观察绿色荧光蛋白的特异性表达。结果:成功构建带有肿瘤特异性survivin启动子驱动的TIEG1基因慢病毒表达载体。感染细胞后,乳腺癌SK-BR-3细胞可观察到绿色荧光表达,而人正常脐静脉内皮细胞基本无表达。半定量RT-PCR证实TIEG1基因在乳腺癌SK-BR-3细胞中过表达。结论:构建的survivin启动子驱动的慢病毒表达载体具有一定的肿瘤特异性,将为实现以慢病毒为载体的TIEG1基因肿瘤靶向治疗提供良好的实验基础。Objective Construction of lentivral vector expressing T1EG1 gene with survivin promoter and identifition of its activity. Methods Survivin promoter and TIEG1 gene full length fragment were obtained by PCR amplification. Lentivial vector containing TIEGI gene and survivin promoter were constructed. The inserted fragment was verified by double enzyme digestion and DNA sequencing. The VSVG pseudotyped lentiviruses were produced and transduced into SK-BR-3 breast cancer cells and normal HUVEC cells. EGFP expression was examined by fluorescence microscopy. Results The lentiviral TIEG1 gene expression vector with survivin promoter was constructed successfu//y and transduced into SK-BR-3 and HUVEC cells. Green fluorescence was observed in SK-BR-3 cells and not in HUVEC cells. Overexpression of TIEG1 gene was verified by RT-PCR in SK-BR-3 cells. Conclusions These data suggest that the survivin promoter drive the specific gene expression in breast cancer cell, which are promising candidates in targeted tumor gene therapy.

关 键 词：乳腺肿瘤 SURVIVIN启动子 TIEG1 慢病毒载体 

分 类 号：R737.9[医药卫生—肿瘤]