[Retroviral vector mediated gene transfer and expression of human GM-CSF in HL-60 leukemic cells].

OBJECTIVE: To study the condition and possibility for introducing foreign gene into myeloid leukemic cells and gene therapy for malignant tumors in hematology. METHODS: With Lipofectin technique, N2A/CMV/hGM-CSF was transfected into packaging cell line PA317 and recombinant retrovirus were obtained. Then the human myeloid leukemic cells HL-60 were infected with recombinant retroviral supernatant. Positive cells were obtained through the neomycin analogue G418 selection. RESULTS: Successful integration of GM-CSF gene into HL-60 cell genome was verified by PCR and Southern Blot, while no specific fragments were amplified by PCR test in HL-60 cells without being transferred or transferred with empty vector N2A. With measurement by GM-CSF dependent cell line TF-1, the level of GM-CSF released by the GM-CSF gene modi-fied HL-60 cell line was 60-200 ng.ml-1.10(-6).24 h-1, while the supernatant of HL-60 cells without being transferred or transferred with empty vector N2A. CONCLUSION: hGM-CSF gene mediated by retroviral vector can achieve efficient transfer and expression in HL-60 leukemic cells.