腺病毒介导KDR启动子的胸苷激酶基因对血管内皮细胞的靶向杀伤作用_医学论文.docVIP

腺病毒介导KDR启动子的胸苷激酶基因对血管内皮细胞的靶向杀伤作用_医学论文 腺病毒介导KDR启动子的胸苷激酶基因对血管内皮细胞的靶向杀伤作用_医学论文 作者：朱欠元，周玉梅，李宝金，毛慕华，彭才圣，郭剑 【关键词】 内皮,血管；KDR启动子；单纯疱疹病毒属；胸苷激酶；腺病毒科 　　Selective killing of vascular endothelial cells based on adenovirusmediated herpes simplex virus thymidine kinase gene transfection under the driving of KDR promoter 　　【Abstract】 AIM: To investigate the target killing effect of adenovirusmediated herpes simplex virus thymidine kinase gene (HSVtk) transfection under the driving of KDR promoter on the vascular endothelial cells in vitro. METHODS：By using AdEasy system, recombinant adenovirus plasmid containing KDR or cytomegalovirus (CMV) promotercontrolled HSVtk gene (AdKDRtk and AdCMVtk) was constructed. After packaging and amplification in 293 cells, the virus was used to infect KDRexpressed human umbilical venous endothelial cells (HUVEC) and KDRunexpressed CNE2. Following administration of ganciclovir (GCV), the survival rate of genetransfected HUVEC and CNE2 was evaluated by using MTT method. RESULTS：The AdEasy System produced a high titer of the recombinant adenovirus (1×1013 pfu/L). Under infection index of 100, with increasing GCV concentration from 0 up to 50 mg/L, the survival rates of AdKDRtktransfected HUVEC and CNE2 decreased from (89.4±4.6)% and (91.5±4.4)% to (22.9±4.7)% and (71.4±2.9)% at proper order respectively (0.01), while the survival rates of AdCMVtktransfected HUVEC and CNE2 declined from (89.9±6.2)% and (90.8±5.7)% to (12.8±2.6)% and (18.8±6.1)%, respectively (Pgt0.05). CONCLUSION：Adenovirusmediated HSVtk transfection under the driving of KDR promoter could yield the specific killing effect on vascular endothelial cells with treatment of GCV. 　　【Keywords】 endothelium, vascular KDR promoter simplex virus thymidine kinase adenoviridac 　　【摘要】 目的：体外实验评估腺病毒介导KDR启动子胸苷激酶系统(HSVtk)杀伤血管内皮细胞的作用. 方法：采用新型AdEasy系统，构建受KDR启动子和巨细胞病毒(CMV)启动子调控并可表达HSVtk基因的AdKDRtk和AdCMVtk，在293细胞中包装、扩增后，体外分别感染表达KDR的人脐静脉血管内皮细胞株（HUVEC）和不表达KDR的人鼻咽癌细胞株CNE2，用丙氧鸟苷(GCV)处理受染细胞，并以MTT法检测其细胞增殖情况. 结果：病毒