A double suicide gene system driven by KDR promoter selectively kills human colon adneocarcinoma SW480 cells / 南方医科大学学报
Journal of Southern Medical University
; (12): 224-227, 2010.
Article
in Zh
| WPRIM
| ID: wpr-269588
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To study the selective killing effect of adenovirus (Ad)-mediated double suicide gene system driven by the KDR promoter (KDR-CDglyTK) on human colon adneocarcinoma SW480 cells.</p><p><b>METHODS</b>KDR-expressing SW480 cells and LS174T cells that did not express KDR were infected by KDR-CDglyTK, and the infection efficiency and the expression of CDglyTK in the cells were detected by RT-PCR. The infected cells were treated with the prodrugs 5-FC and GCV at different concentrations, and the cell-killing effects and bystander effects were evaluated by MTT method. DNA content and the cell cycle changes in SW480 cells were detected by flow cytometry.</p><p><b>RESULTS</b>The expression of green fluorescent protein (GFP) was observed in 95% of the infected SW480 and LS174T cells with a multiplicity of infection (MOI) of 100. RT- PCR demonstrated that the product of CD/TK gene existed in SW480 cells infected by Ad- KDR- CD/TK, but not in infected LS174 cells. The infected SW480 cells exhibited high sensitivity to the prodrugs, but the infected LS174T cells did not (P<0.01). Bystander effects of the double suicide gene system were observed in the coculture of the infected and non-infected SW480 cells. At the MOI of 100, treatment of the infected cells with the prodrugs resulted in increased cell percentage in G(0)-G(1) phase and decreased percentage in S phase and the prodrug-treated cells showed an apoptotic peak in flow cytometry.</p><p><b>CONCLUSION</b>CDglyTK fusion gene system driven by the KDR promoter selectively kills and induces the apoptosis of the KDR-CDglyTK SW480 cells.</p>
Full text:
1
Index:
WPRIM
Main subject:
Pathology
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Thymidine Kinase
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Recombinant Fusion Proteins
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Genetic Therapy
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Adenocarcinoma
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Adenoviridae
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Promoter Regions, Genetic
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Apoptosis
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Colonic Neoplasms
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Vascular Endothelial Growth Factor Receptor-2
Limits:
Humans
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2010
Type:
Article