Serum factors regulate 5-lipoxygenase activity in maturating HL60 cells.
5-Lipoxygenase activity in DMSO-differentiated HL60 cells is regulated by human serum. The serum effect depended on the differentiation state of the cells. For a stimulatory effect to occur, it was required that the cells had been treated with DMSO before addition of serum. After this regimen, the HL60 cells acquired the same high 5-LO activity as found for human neutrophils isolated from peripheral blood (about 9-times higher than for HL60 cells treated only with DMSO). On the other hand, when serum was added together with DMSO and present during the entire differentiation period (seven days), or withdrawn after the first four days, the 5-LO activity did not increase. 5-LO activity of HL60 cells covaried with the expression of the CD14 molecule, a marker for myeloid cell maturation which was recently identified as a receptor for the complex of LPS and LPS-binding protein. These serum effects on 5-LO activity were only observed for intact cells. The prominent increase in 5-LO activity induced by serum was not concomitant with similar changes in the expression of 5-LO or 5-LO-activating protein (FLAP), as judged from analyses of immunoreactive protein and mRNA. Also, the high 5-LO activity induced by serum was rather insensitive to the drug MK886 under our standard assay conditions, which included addition of exogenous arachidonic acid (40 microM). The results indicate that additional cellular components of importance for 5-LO activity in HL60 cells become operative after serum treatment, and that mere expression of 5-LO and FLAP is insufficient for high 5-LO activity in intact cells.
Steinhilber
D
D
Department of Physiological Chemistry, Karolinska Institutet, Stockholm, Sweden.
Hoshiko
S
S
Grunewald
J
J
Rådmark
O
O
Samuelsson
B
B
D001094:Arachidonate 5-Lipoxygenase / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D002462:Cell Membrane; D004790:Enzyme Induction; D006098:Granulocytes / Q000378:metabolism*; D006801:Humans; D007211:Indoles; D015289:Leukotrienes / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000032:analysis; D014407:Tumor Cells, Cultured / Q000378:metabolism; D002722:Chlorobenzenes
D001094:Arachidonate 5-Lipoxygenase; D007211:Indoles; D015289:Leukotrienes; D012333:RNA, Messenger; C060893:MK-886; D002722:Chlorobenzenes