Failure of F-Met-Leu-Phe to induce chemotaxis in differentiated promyelocytic (HL-60) leukemia cells.
Treatment of HL-60 promyelocytic leukemia cells with dibutyryl cyclic adenosine 3',5'-monophosphate (dbcAMP) induced these cells to differentiate toward mature neutrophilic granulocytes (PMN). DbcAMP was found to produce a rapid time- and dose-dependent inhibition of HL-60 cell proliferation, reaching a maximum after 48 hr treatment of the cells with 250-500 microM. This was associated with morphologic alterations consistent with maturing PMN. Using immunofluorescence and flow cytometry, we found that dbcAMP-treated HL-60 cells expressed some, but not all, surface markers characteristic of mature phagocytes. Thus receptors for the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP); the monocyte/granulocyte markers My-8, Mo-1, and Leu-15; as well as the monocyte markers Mo-2 and My-4 were identified on the cells. In contrast, dbcAMP-treated cells did not express the PMN-specific antigen DL1.2. We also found that dbcAMP-treated HL-60 cells produced hydrogen peroxide in response to the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA). However, the magnitude of this response was significantly less than that observed in mature PMN. In contrast, fMLP-stimulated levels of hydrogen peroxide production were comparable in both cell types. In addition, despite the fact that dbcAMP-treated cells expressed fMLP receptors, they did not exhibit directed migration toward this chemotactic peptide. Taken together, these data suggest that dbcAMP-induced differentiation of HL-60 cells is incomplete.
Sirak
A A
AA
Department of Pharmacology, Rutgers University, Piscataway, NJ 08855-0789.
Laskin
J D
JD
Robertson
F M
FM
Laskin
D L
DL
D000954:Antigens, Surface / Q000032:analysis; D003994:Bucladesine / Q000494:pharmacology; D002087:Butyrates / Q000494:pharmacology; D002454:Cell Differentiation; D002455:Cell Division; D002634:Chemotaxis, Leukocyte / Q000187:drug effects*; D005576:Colforsin / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D015473:Leukemia, Promyelocytic, Acute / Q000276:immunology*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D002087:Butyrates; D005576:Colforsin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D003994:Bucladesine; D006861:Hydrogen Peroxide; D015232:Dinoprostone