转录靶向性KDR启动子调控双自杀基因治疗肺癌的实验研究  

Experimental Study on the Transcriptional Targeting Gene Therapy of Double Suicide Gene Driven by KDR Promotor in Lung Cancer

作　　者：车国卫[1] 周清华[1] 刘伦旭[1] 步宏[2] 王艳萍[3] 朱文[3] 陈晓禾[3] 唐小军 

机构地区：[1]四川大学华西医院胸心血管外科,成都610041 [2]四川大学华西医院病理科,成都610041 [3]四川大学华西医院四川省肺癌分子重点实验室,成都610041

出　　处：《科技导报》2006年第5期28-31,共4页Science & Technology Review

基　　金：国家自然科学基金项目(30400199);中国博士后科学基金项目(2004035697)

摘　　要：从人肺癌细胞株中克隆KDR基因的启动子(kinasedomainreceptorpromotor,KDRp),构建KDR基因启动子调控的双自杀基因(CDglyTK)真核表达质粒pcDNA3-KDRp-CdglyTK,将其导入ECV304、L9981和NL9980细胞,建立相应的转基因细胞系,并应用不同的前药处理。体内、外实验结果显示:KDR启动子调控的双自杀基因在KDR高表达人肺癌细胞和人脐静脉内皮细胞靶向表达,而在KDR不表达的正常细胞或正常血管内皮细胞中未检测到双自杀基因表达;联合应用5-FC和GCV处理,对转双自杀基因细胞的杀伤作用显著高于单独应用5-FC或GCV,且二者显示了良好的药物协同作用。Human kinase domain receptor promoter （KDRp） has been cloned into the pcDNA3 -CDglyTK to construct the pcDNA3-KDRp- CDglyTK specialized expression vector. The pcDNA3-KDRp-CDglyTK plasmid was transfected into ECV304, L9981 and NL9980 cell, and established ECV304-cdtk, L9981-cdtk and NL9980-cdtk cell lines. The above cell lines were treated with 5-FC and/or GCV respectively. The result suggest that cdtk gene was specialized expressed in ECV304-cdtk, L9981-cdtk and NL9980-cdtk KDR-positive cells with KDR gene high expression, and prodrug/KDRp-CDglyTK system was effective in killing effect in KDR-positive cells. The killing effect in combined drug （5-FC and GCV） was remarkable stronger than that in GCV alone and 5-FC alone in the same pre-drug dose, and a significantly synergetic cytotoxic effect was found in vivo and in vitro.

关 键 词：转录靶向性 基因治疗 双自杀基因 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]