[Killing effect of human pulmonary adenocarcinoma cells with TK + CD/5-Fc + GCV coexpression suicide gene systems].
Cao Hq, Yongxiang Zhao, Meng Xm, Ding Jf
Abstract
Cao Hq, Yongxiang Zhao, Meng Xm, Ding Jf
Abstract
OBJECTIVE: To investigate the different killing effect to human pulmonary adenocarcinoma cell line cells GLC-82 with coexpressed double suicide genes compared with single gene. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
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OBJECTIVE: To investigate the different killing effect to human pulmonary adenocarcinoma cell line cells GLC-82 with coexpressed double suicide genes compared with single gene. METHODS: 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. RESULTS: 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. CONCLUSION: 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.
Key concepts: Suicide gene, Molecular biology, Cytosine deaminase, Transfection, Lipofectamine, Thymidine kinase, Cytotoxic T cell, Cell culture