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  

Selective killing of vascular endothelial cells based on adenovirus-mediated herpes simplex virus thymidine kinase gene transfection under the driving of KDR promoter

作　　者：朱欠元[1] 周玉梅[1] 李宝金[2] 毛慕华[1] 彭才圣[1] 郭剑[1] 

机构地区：[1]井冈山学院医学院耳鼻喉科,江西吉安343000 [2]北京大学深圳医院肝胆胰外科,广东深圳518036

出　　处：《第四军医大学学报》2006年第22期2036-2038,共3页Journal of the Fourth Military Medical University

基　　金：江西省卫生厅科技资助(20052041)

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(HSV-tk)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HSV-tk基因的AdKDR-tk和AdCMV-tk,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CNE-2,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:病毒滴度均为1×1013pfu/L.在感染复数(MO I)为100的条件下,GCV浓度由0增至50 mg/L时感染含AdKDR-tk的HUVEC细胞和CNE-2细胞存活率从(89.4±4.6)%和(91.5±4.4)%分别下降至(22.9±4.7)%和(71.4±2.9)%(P<0.01),而感染AdCMV-tk的HUVEC细胞和CNE-2细胞存活率从(89.9±6.2)%和(90.8±5.7)%分别下降至(12.8±2.6)%和(18.8±6.1)%(P>0.05).结论:腺病毒介导KDR启动子-胸苷激酶系统具有特异性杀伤血管内皮细胞作用.AIM： To investigate the target killing effect of adenovirus-mediated herpes simplex virus thymidine kinase gene （HSV-tk） transfeetion under the driving of KDR promoter on the vascular endothelial cells in vitro. METHODS：By using AdEasy system, recombinant adenovirus plasmid containing KDR or eytomegalovirus （CMV） promoter-controlled HSV-tk gene （AdKDRtk and AdCMV-tk） was constructed. After packaging and amplification in 293 cells, the virus was used to infect KDR-expressed human umbilical venous endothelial cells （ HUVEC ） and KDR- unexpressed CNE-2. Following administration of ganeielovir （GCV） , the survival rate of gene-transfected HUVEC and CNE-2 was evaluated by using MTr method. RESULTS：The AdEasy System produced a high titer of the recombinant adenovirus （ 1 × 10^13 pfu/L）. Under infection index of 100, with increasing GCV concentration from 0 up to 50 mg/L, the survival rates of AdKDR- tk-transfected HUVEC and CNE-2 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 （P 〈0. 01）, while the survival rates of AdCMV-tk-transfected HUVEC and CNE-2 declined from （89.9±6.2）% and （90.8±5.7）% to （12.8± 2.6）% and （18. 8 ± 6. 1）%, respectively （P 〉 0. 05）. CONCLUSION ：Adenovirus-mediated HSV-tk transfection under the driving of KDR promoter could yield the specific killing effect on vascular endothelial cells with treatment of GCV.

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 

分 类 号：R739.63[医药卫生—肿瘤]