[Targeted killing of colorectal tumor cells by lentiviral constructs containing CD/TK suicide genes and KDR promoter].
OBJECTIVE
To investigate the selective killing of colorectal tumor cells by lentivirus-mediated double suicide gene under the regulation of KDR promoter.


METHODS
293T packaging cells were transfected with the plasmid FGW-KDRP-CD/TK to obtain the infectious viruses. KDR-expressing LoVo cells and LS174T cells that did not produce KDR were transfected with the recombinant virus, and the transfection efficiency was evaluated by the fluorecence microscope. RT-PCR was employed to examine the expression of CDglyTK. After treatment of the cells with 5-FC and GCV, the killing effects on the two cell lines were evaluated.


RESULTS
The recombinant construct showed similar infection rate of the two cell lines. RT-PCR demonstrated that CDglyTK gene was expressed only in LoVo cells infected with FGW-KDRP-CD/TK but not in LS147T cells, and the sensitivity of the two cell lines to the prodrugs was significantly different (P<0.001). The killing effect of the double suicide gene was much stronger than that of single suicide gene administered (P<0.001).


CONCLUSIONS
The double suicide gene driven by KDR promoter has specific killing effect on the KDR-expressing colorectal tumor cells.
Chen
Hai-jin
HJ
Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. chenhaijin@sina.com
Huang
Zong-hai
ZH
Wu
Ai-guo
AG
Yu
Jin-long
JL
Su
Guo-qiang
GQ
D000963:Antimetabolites / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D043525:Cytosine Deaminase / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry; D005437:Flucytosine / Q000494:pharmacology; D015774:Ganciclovir / Q000494:pharmacology; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics
D000963:Antimetabolites; D011993:Recombinant Fusion Proteins; D005437:Flucytosine; D013937:Thymidine Kinase; D040301:Vascular Endothelial Growth Factor Receptor-2; D043525:Cytosine Deaminase; D015774:Ganciclovir