摘要
目的 通过细菌内同源重组 ,制备表达全长smac基因的腺病毒 ,初步观察其体外对肿瘤细胞的杀伤作用。方法 构建含smac基因的腺病毒穿梭载体pAdTrack smac ,与骨架载体 pAdEasy1在细菌内重组为pAd .smac,经 2 93细胞包装为增殖缺陷性腺病毒。体外感染各种肿瘤细胞 ,用苔盼蓝拒染法观察其对肿瘤细胞的杀伤作用。结果 利用AdEasyTM 系统成功构建了Ad .smac及对照腺病毒Ad .GFP ,多种肿瘤细胞感染Ad .smac后均出现细胞凋亡现象 ,对肿瘤细胞的杀伤率达 30 %以上。结论 利用细菌内质粒同源重组法可快速简捷地制备表达外源基因的腺病毒 ,Ad .smac在体外通过诱导细胞凋亡而有效地杀伤肿瘤细胞 ,为Smac在肿瘤治疗的应用提供了一定的试验参考依据。
Objective To generate adenovirus Ad.to smac, express the full-length smac cDNA by means of homologous recombination in bacteria. Also explore the anti-tumor effect of Ad.smac in vitro. Methods Construct shuttle plasmid pAdeasy1 in bacteria was performed to generate pAd.smac. Replication deficient adenovirus was packed and propagated in the cell 293. The anti-tumor effects of smac on various tumor cells were observed after Ad.smac infection,by using trypan blue exclusion method. Results Ad.smac and control adenovirus Ad. GFP were successfully constructed by AdEasy TM System. Tumor cells proceeded apoptosis after Ad.smac infection and over 30% cells were killed. Conclusion Homologous recombination in bacteria is a simple and convenient strategy to genarate adenovirus expressing exogenous genes. Ad.smac can kill tumor cells effectively in vitro through induction of apoptosis, and can be used as a potential weapon for tumor therapy.
出处
《肿瘤》
CAS
CSCD
北大核心
2003年第6期486-489,共4页
Tumor
基金
国家自然科学基金资助项目 (编号 :3 0 0 0 0 0 0 8)
