[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.


CONCLUSIONS
hGM-CSF gene mediated by retroviral vector can achieve efficient transfer and expression in HL-60 leukemic cells.
Institute of Hematology, Tongji Medical University, Union Hospital, Wuhan.
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D018014:Gene Transfer Techniques; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis / Q000235:genetics*; D018922:HL-60 Cells / Q000378:metabolism; D006801:Humans; D011994:Recombinant Proteins; D012190:Retroviridae / Q000235:genetics*
D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor