构建以hTERT、Cox-2启动子调控的肿瘤特异增殖腺病毒  

Construction of hTERT and Cox-2 dependent oncolytic adenovirus

作　　者：华汉巨[1] 毛晨宇[1] 滕理送[1] 

机构地区：[1]浙江大学医学院附属第一医院肛肠外科,浙江杭州310003

出　　处：《实用肿瘤杂志》2006年第5期417-420,共4页Journal of Practical Oncology

基　　金：国家自然科学基金(30572155);浙江省医药卫生重点科技项目(2004ZD004)资助

摘　　要：目的应用同源重组的方法构建以hTERT和Cox-2启动子调控增殖的肿瘤特异性增殖腺病毒。方法将hTERT和Cox-2启动子从人类白细胞基因组中亚克隆出来,并将启动子分别插入到腺病毒穿梭载体pd306上的E 1A和E 1B基因前,使hTERT和Cox-2启动子分别调控腺病毒必须基因E 1A和E 1B的表达,再将构建后的pd306和腺病毒的骨架质粒BHGE 3在A d293细胞内进行同源重组,并用重组后的病毒感染H e la细胞检测病毒对肿瘤细胞的杀伤力。结果成功构建了hTERT和Cox-2启动子,并将两个启动子连接入腺病毒载体,产生了具有感染力的可增殖腺病毒。结论经hTERT和Cox-2启动子调控增殖的肿瘤特异性增殖腺病毒对H e la细胞具有杀伤力,为进一步研究病毒在体内、外对各种肿瘤细胞的特异性杀伤力、安全性奠定了基础。Objective To construct a hTERT and Cox-2 promoter regulated oncolytic virus by replacing the endogenous adenovirus Ela and Elb promoter,respectively. Methods The promoter of hTERT and Cox-2 was subcloned from human genomic DNA, then ligated to shuttle vector pd306 which contains the Ela and Elb gene of adenovirus to control the expression of Ela and Elb,respectively. The constructed shuttle vector was cotransfected with adenovirus hone vector in Ad293 cell to gain oncolytic adenovirus and determine its titer. Oncolytic virus was used to infect Hela cells to detect its execution to tumor cells. Result （1）The promoter of hTERT and Cox-2 was subcloned. （2）The hTERT and Cox-2 promoter controlled oncolytic adenovirus was gained. （3）The gained virus had ability to kill Hela cells. Contrutions The hTERT and Cox-2 dependent oncolytic adenoirus has been constructed for further research of effection of oncolytic virus on tumor cells and normal cells.

关 键 词：肿瘤 腺病毒 人 前列腺素内过氧化物合酶 启动区(遗传学) 端粒 基因 

分 类 号：R73-3[医药卫生—肿瘤]