Cao H Q, Zhao Y, Meng X M, Ding J F
Molecular Medicine Center, FuWai Heart Institute, FuWai Heart Hospital, CAMS, PUMC, Beijing 100037, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2001 Apr;23(2):168-72.
To investigate the different killing effect to human pulmonary adenocarcinoma cell line cells GLC-82 with coexpressed double suicide genes compared with single gene.
Recombinant expression vectors containing CD (cytosine deaminase) and/or TK (thymidine kinase) gene under CMV promoter were constructed successfully. The vectors were transfected to GLC-82 tumor cell lines by use of lipofectamine. The clones were picked out after G418 selection. Extraneous gene integration and expression were confirmed by PCR and semi-quantitative RT-PCR. The cytotoxicity to these transgenic cells under treatment with 5-Fc and GCV were measured by MTT assays.
Double and single suicide gene transfer were both stably expressed in GLC-82 cells. The cytotoxic effects of co-expressed TK-CD genes were superior than that of the single gene.
The CD + TK/5-Fc + GCV co-expression system is more effective for killing effect of tumor cells than CD/5-Fc or TK/GCV system alone.
探讨共表达双自杀基因与单基因对人肺腺癌细胞系GLC-82的杀伤效果差异。
成功构建了在巨细胞病毒（CMV）启动子调控下含胞嘧啶脱氨酶（CD）和/或胸苷激酶（TK）基因的重组表达载体。利用脂质体将载体转染至GLC-82肿瘤细胞系。经G418筛选后挑出克隆。通过聚合酶链反应（PCR）和半定量逆转录聚合酶链反应（RT-PCR）证实外源基因的整合与表达。采用噻唑蓝（MTT）法检测5-氟胞嘧啶（5-Fc）和丙氧鸟苷（GCV）处理下这些转基因细胞的细胞毒性。
双自杀基因和单自杀基因转染在GLC-82细胞中均稳定表达。共表达TK-CD基因的细胞毒性作用优于单基因。
与单独的CD/5-Fc或TK/GCV系统相比，CD+TK/5-Fc+GCV共表达系统对肿瘤细胞的杀伤效果更有效。
