Lipoxygenase metabolism is required for interleukin-3 dependent proliferation and cell cycle progression of the human M-07e cell line.
The cell line M-07e requires either Interleukin-3 (IL-3) or granulocyte-macrophage colony stimulating factor (GM-CSF) for proliferation in vitro. Cells deprived of growth factor for up to 48 hours remain viable but no longer divide. The growth-factor-deprived M-07e cells begin to divide within 48 hours of reexposure to IL-3. Flow cytometric analysis of M-07e cells labeled with hypotonic propidium iodide demonstrates that the percentage of cells undergoing DNA synthesis decreases from 24%, in a log phase population of IL-3 stimulated cells, to 1% when cells are deprived of IL-3 for 24 hours. IL-3-deprived cells accumulate predominantly in a flow cytometry peak representative of G0/G1. DNA synthetic activity, as determined by tritiated thymidine uptake and flow cytometry, resumes between 12 and 18 hours after reexposure to IL-3, reaching a peak of up to 40% by 24 hours and returning to log phase levels by 72 hours. Prior to initiation of DNA synthesis, increases are seen in mRNA levels for five-lipoxygenase-activating protein (FLAP). Following reexposure to IL-3, a rapid time-dependent biosynthesis of leukotriene D4 (LTD4) is induced by M-07e cells. When IL-3 is added in the presence of any of three lipoxygenase inhibitors tested (Piriprost, caffeic acid, nordihydroguiaretic acid) or FLAP inhibitor, MK-886, there is dose-dependent inhibition of the resumption of proliferation and of DNA synthesis. Flow cytometric cell cycle analysis demonstrates that the inhibited cells remain in the G0/G1 population and do not progress through the cell cycle. These results are consistent with our previous observation that an intact lipoxygenase pathway is necessary for hematopoietic growth-factor-stimulated colony formation of normal bone marrow myeloid progenitors and suggest that the induction of a lipoxygenase metabolite or metabolites is necessary for myeloid cells to progress through the cell cycle when stimulated by a hematopoietic growth factor.
Miller
A M
AM
Tulane Cancer Center, Tulane University, New Orleans, Louisiana 70112, USA.
Allen
B S
BS
Ziboh
V
V
D058927:5-Lipoxygenase-Activating Proteins; D001094:Arachidonate 5-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D001854:Bone Marrow Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002460:Cell Line / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D005434:Flow Cytometry; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology*; D015289:Leukotrienes / Q000096:biosynthesis / Q000378:metabolism; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D013936:Thymidine / Q000493:pharmacokinetics; D014316:Tritium / Q000493:pharmacokinetics
D058927:5-Lipoxygenase-Activating Proteins; C548592:ALOX5AP protein, human; D002352:Carrier Proteins; D007377:Interleukin-3; D015289:Leukotrienes; D016859:Lipoxygenase Inhibitors; D008565:Membrane Proteins; D012333:RNA, Messenger; D014316:Tritium; D017228:Hepatocyte Growth Factor; D001094:Arachidonate 5-Lipoxygenase; D013936:Thymidine