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Cytotoxicity and bystander effect of lentiviral vector-mediated YCD/5-FC suicide gene system against leukemia Jurkat cells
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Abstract:
Objective: To investigate the cytotoxicity and bystander effects of lentiviral vector-mediated yeast cytosine deaminase (YCD) suicide gene against the human leukemia Jurkat cells. Methods:YCD-GFP lentiviral vector was prepared and transfected into Jurkat cells (Jurkat/YCD-GFP cells). The expression of YCD-GFP gene was examined by flow cytometry. The cytotoxicity and bystander effects of pro-drug 5-fluorocytosine (5-FC) on Jurkat/YCD-GFP cells were measured in vitro. Nude mice bearing-Jurkat/YCD-GFP cell tumors were established and the cytotoxicity and bystander effects of 5-FC on tumor cells were measured in vivo. Results: YCD-GFP lentiviral vector and Jurkat/YCD-GFP cells (Jurkat cells infected by YCD-GFP lentiviral vector) were successfully prepared; the infection rate of YCD-GFP lentiviral vector on Jurkat cells was 96.93%. 5-FC (232 μmol/L for 72h) killed (95.61±2.07)% of Jurkat/YCD-GFP cells, and 5-FC level in the culture supernatant decreased by about 80%. Bystander effect of Jurkat/YCD-GFP cells could be observed both in mixed culture (effect∶target=1∶10, \[68.69± 4.97\]% vs \[97.87±1.11\]%, P<0.05) and in Transwell culture (effect∶target=1∶25, \[1.46±0.27\]×105 vs \[3.00±0.16\]×105，P<0.01) . In vivo experiment results demonstrated that YCD/5-FC suicide gene system had strong cytotoxicity and bystander effects in Jurkat/YCD-GFP tumor bearing mice (P<0.05). Conclusion: The lentiviral vector-mediated YCD/5-FC suicide gene system has marked cytotoxicity and bystander effects against human leukemia Jurkat cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China （No. 30871100，No.30172347）, and the Key Foundation of Science and Technology Commission of Shanghai (No.8JC1406500)
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