Induction of the fms proto-oncogene product in HL-60 cells by vitamin D: a flow cytometric analysis.
Agents which induce monocytic characteristics in HL-60 human acute promyelocytic leukemia cells induce mRNA for the fms proto-oncogene, which encodes the receptor for M-CSF. Previous studies of fms expression in HL-60 cells have characterized chiefly induction by phorbol esters of fms mRNA. Our studies of fms expression in HI-60 cells have characterized induction by vitamin D3 of the fms protein. We have used flow cytometry to correlate fms antigen with a monocyte-specific differentiation antigen recognized by antibody MO2 (CD14), with DNA content, and with the nuclear antigen Ki-67, a marker of cell cycling. HL-60 cells were cultured with or without 1 microM vitamin D for 7 days. fms antigen was found on 42 +/- 5.8% of the cells cultured without vitamin D, but on 63 +/- 4.3% of the cells cultured with vitamin D. MO2 binding was detected on only 2 +/- 0.5% of the cells without vitamin D, but on 59 +/- 9% with vitamin D. Cells cultured with vitamin D that were fms-positive were also predominantly (83%) MO2-positive. Analysis of DNA content, measured by propidium iodide staining, showed that 57 +/- 1.5% of cells cultured without vitamin D, but 93 +/- 0.5% of cells cultured with vitamin D, were in the G0/G1 cell cycle phase. Analysis of nuclear antigen Ki-67 revealed that, of the vitamin D-treated cells that were fms-positive, a significant proportion (37%) were still cycling. We conclude that (1) fms is demonstrable on some uninduced HL-60 cells, (2) when HL-60 cells are induced to develop monocytic characteristics by vitamin D, fms induction is part of the program for monocytic differentiation that includes MO2 expression, yet (3) some induced cells expressing fms are still cycling.
Rowley
P T
PT
Department of Medicine, University of Rochester School of Medicine, New York 14642.
Farley
B
B
Giuliano
R
R
LaBella
S
S
Leary
J F
JF
D000943:Antigens, Differentiation / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002762:Cholecalciferol / Q000494:pharmacology*; D004273:DNA, Neoplasm / Q000378:metabolism; D005434:Flow Cytometry; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects*; D016236:Genes, fms / Q000187:drug effects*; D006801:Humans; D019394:Ki-67 Antigen; D015473:Leukemia, Promyelocytic, Acute / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D016281:Oncogene Protein gp140(v-fms) / Q000378:metabolism*; D000090063:Proto-Oncogene Mas; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000276:immunology / Q000378:metabolism
D000943:Antigens, Differentiation; D004273:DNA, Neoplasm; D019394:Ki-67 Antigen; C000715470:MAS1 protein, human; D009687:Nuclear Proteins; D016281:Oncogene Protein gp140(v-fms); D000090063:Proto-Oncogene Mas; C036763:monocyte-macrophage differentiation antigen; D002762:Cholecalciferol