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[Selective cytotoxic effect of lentivirus-mediated double suicide gene transfer on human gastric adneocarcinoma cells].
- Source :
-
Nan fang yi ke da xue xue bao = Journal of Southern Medical University [Nan Fang Yi Ke Da Xue Xue Bao] 2010 Jan; Vol. 30 (1), pp. 47-50. - Publication Year :
- 2010
-
Abstract
- Objective: To study the selective cytotoxic effect of lentivirus-mediated double suicide gene (CD/TK) against human gastric carcinoma cells SGC-7901 in vitro.<br />Methods: SGC-7901 cells were infected with FGW-KDRP-CD/TK vector and the infection efficiency was observed under a fluorescence microscope. The morphological changes of the infected cells were observed by Giemsa staining. Flow cytometry (FCM) was employed for cell cycle analysis, and the expression of CD/TK was detected by RT-PCR. The infected cells were then treated with the prodrugs ganciclovir (GCV) and/or 5-fluorocytosine (5-FC) at different concentrations, and the cytotoxic effects were evaluated using MTT method.<br />Results: The infection efficiency of the lentiviral vector in SGC-7901 cells increased with the titer of the virus, which produced no significant effect on the cancer cell morphology in vitro or on the percentages of G0-G1, G2-M and S phase cells (P>0.05). RT-PCR demonstrated the expression of CD/TK gene in SGC-7901 cells infected by FGW-KDRP-CD/TK. The infected cells were highly sensitive to the prodrugs with a dose-dependent cytotoxic effect within a specific concentration range of the drugs, whereas the non-infected cells were not sensitive to the prodrugs. Combined use of the two prodrugs produced an obviously stronger inhibitory effect than either of the them (P<0.05). When combined, GCV and 5-FC at the concentration of 0.1+40, 1+80, 10+160, and 100+320 mg/L demonstrated a synergetic effect with a CDI<1.<br />Conclusion: Lentivirus-mediated CD/TK fusion gene system can selectively kill gastric cancer cells, and the two prodrugs show a synergistic cytotoxic effect.
- Subjects :
- Adenocarcinoma genetics
Adenocarcinoma pathology
Cell Line, Tumor
Cytosine Deaminase biosynthesis
Cytotoxins pharmacology
Genetic Therapy
Genetic Vectors genetics
Humans
Lentivirus metabolism
Recombinant Fusion Proteins biosynthesis
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins pharmacology
Stomach Neoplasms genetics
Thymidine Kinase biosynthesis
Vascular Endothelial Growth Factor Receptor-2 genetics
Vascular Endothelial Growth Factor Receptor-2 metabolism
Cytosine Deaminase genetics
Genes, Transgenic, Suicide genetics
Lentivirus genetics
Stomach Neoplasms pathology
Thymidine Kinase genetics
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1673-4254
- Volume :
- 30
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nan fang yi ke da xue xue bao = Journal of Southern Medical University
- Publication Type :
- Academic Journal
- Accession number :
- 20117982