Arachidonic acid metabolism by a vitamin D3-differentiated human leukemic cell line.
HL-60 is a human promyelocytic cell line that differentiates along the granulocytic pathway when incubated with dimethylsulfoxide and along the monocytic pathway when incubated with 1,25-(OH)2D3. We compared arachidonic acid metabolism in undifferentiated, DMSO-differentiated, and 1,25-(OH)2D3-differentiated cells. DMSO- and 1,25-(OH)2D3-differentiated cells metabolized exogenous arachidonic acid to both cyclo-oxygenase products (predominantly thromboxane B2 and prostaglandin E2) and 5-lipoxygenase products, including leukotriene B4. Undifferentiated cells produce these metabolites in much smaller amounts. DMSO-differentiated cells released a large percentage of phospholipid-bound arachidonic acid in response to stimulation with the ionophore A23187, zymosan, or formylmethionylleucylphenylalanine (FMLP). DMSO-differentiated cells stimulated with A23187 converted released arachidonate to LTB4 and TxB2. In contrast, 1,25-(OH)2D3-differentiated cells released a smaller percentage of phospholipid-bound arachidonate in response to stimuli, and undifferentiated cells released none at all. The three cell types (undifferentiated, DMSO-differentiated, and 1,25-(OH)2D3-differentiated) were homogenized and the 10,000 X g supernatant incubated with [14C]arachidonic acid. The supernatants from the homogenates of the DMSO- and 1,25-(OH)2D3-differentiated cells metabolized [14C]arachidonic acid to cyclooxygenase and lipoxygenase products, but the supernatant from the homogenate of undifferentiated cells did not. These data indicate that differentiation of HL-60 cells with DMSO or 1,25-(OH)2D3 induces cyclooxygenase and 5-lipoxygenase and induces mechanisms for the release of arachidonate from phospholipids by soluble and particulate stimuli.
Stenson
W F
WF
Department of Medicine, Jewish Hospital, Washington University Medical Center, St. Louis, MO.
Teitelbaum
S L
SL
Bar-Shavit
Z
Z
D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002458:Cell Fractionation; D004121:Dimethyl Sulfoxide; D006801:Humans; D007938:Leukemia / Q000378:metabolism*; D007975:Leukotriene B4 / Q000096:biosynthesis; D008055:Lipids / Q000032:analysis; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D011863:Radioimmunoassay; D013929:Thromboxane B2 / Q000096:biosynthesis; D014407:Tumor Cells, Cultured; D015054:Zymosan / Q000494:pharmacology
D001095:Arachidonic Acids; D008055:Lipids; D007975:Leukotriene B4; D016718:Arachidonic Acid; D000001:Calcimycin; D013929:Thromboxane B2; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015054:Zymosan; D002117:Calcitriol; D004121:Dimethyl Sulfoxide