摘要
目的 观察逆转录病毒介导的双自杀基因对淋巴瘤细胞的杀伤作用 ,探讨淋巴瘤的基因治疗方法。方法 在脂质体的介导下将含有双自杀基因的逆转录病毒载体 pWZLneoCDglytk导入包装细胞PA317,经G418筛选后大量培养产病毒的阳性克隆PA 317/CD +tk细胞株 ,收集病毒上清 ,浓缩后转染Raji淋巴瘤细胞 ,再次经G 418筛选 ,获得稳定表达双自杀基因的Raji/CD +tk细胞株。采用RT -PCR法检测双自杀基因的表达。MTT法测定转基因组及未转基因组Raji细胞的存活率。 结果 双自杀基因在Raji细胞中可稳定表达 ,联合使用 5 -氟胞嘧啶 ( 5 -FC)和无环鸟苷 (GCV )对细胞增殖的杀伤作用及旁杀伤效应高于单独使用 5 -FC或GCV。
Objective To investigate the killing effect of cytosine deaminase(CD) and herpes simplex virus thymidine kinase(HSV-tk) double suicide genes on lymphoma Raji cells mediated by retrovirus compared with that of single gene.Methods CD and HSV-tk double suicide genes were transfected into PA317 cells using lipofectamine.The positive clones were picked out and cultured after G418 selection.The viral supernatant was collected and concentrated.The lymphoma cells were infected with the concentrated virus containing the double suicide genes.After G418 selection,RT-PCR was used to demonstrate the successful transcription of CD and HSV- tk genes.The Raji/CD+tk and Raji cells in culture were respectively treated with 5-FC,GCV or both.The cytoxicity efficiency was evaluated by MTT method.Results Double suicide genes were stably expressed in Raji lymphoma cells after being infected with virus.The killing effect of Combined 5-FC with GCV on Raji/CD+tk cells was more significant than that of 5-FC or GCV alone.Conclusion The CD+tk/5FC+GCV co-expression system is more effective for killing lymphoma cells than the CD/5FC tk/GCV system.
出处
《实用癌症杂志》
2003年第3期228-230,共3页
The Practical Journal of Cancer
基金
国家自然科学基金资助 (No :30 0 70 32 1 )
