title: Effects of lipopolysaccharide on phospholipase A2 activity and tumor necrosis factor expression in HL-60 cells.
abstract: LPS has been identified as a potent activator of mononuclear phagocytes. This activation is associated with TNF gene expression. The intracellular signaling mechanisms responsible for this effect, however, are unknown. The present studies demonstrate that LPS induces TNF transcripts in HL-60 promyelocytic leukemia cells. Because previous studies have demonstrated that eicosanoids are involved in the regulation of TNF gene expression in these cells, we examined the effects of LPS on activation of the arachidonic acid cascade. The results demonstrate that LPS stimulates phospholipase A2 activity and the hydrolysis of both 1,2-dipalmitoyl phosphatidylcholine and 1-steroyl 2-arachidonoyl phosphatidylcholine. In contrast, there was no detectable effect of LPS on activation of protein kinase C. We also demonstrate that inhibition of phospholipase A2 activity with bromophenacyl bromide or quinacrine blocks the induction of TNF transcripts by LPS. These findings suggested that LPS induces TNF gene expression through formation of arachidonic acid metabolites. Indeed, similar results were obtained with mellitin, a known activator of phospholipase A2 and eicosanoid production. Previous studies have also suggested that TNF mRNA levels are increased in HL-60 cells by the 5-lipoxygenase pathway and, in the present work inhibitors of this enzyme blocked LPS-induced TNF expression. Moreover, the cyclooxygenase metabolite, PGE2, as well as dibutyryl cAMP, inhibited the induction of TNF transcripts by LPS. Taken together, these results suggest that LPS induces TNF gene expression through activation of phospholipase A2 and that the level of this induction is regulated by activity of the 5-lipoxygenase and cyclooxygenase pathways.
authors.affiliation: Laboratory of Clinical Pharmacology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115.
authors.forename: M
authors.initials: M
authors.lastname: Mohri
authors.forename: D R
authors.initials: DR
authors.lastname: Spriggs
authors.forename: D
authors.initials: D
authors.lastname: Kufe
mesh_terms: D000098:Acetophenones / Q000494:pharmacology; D003994:Bucladesine / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D007654:Ketoconazole / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D009637:Masoprocol / Q000494:pharmacology; D008555:Melitten / Q000494:pharmacology; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011493:Protein Kinase C / Q000378:metabolism; D011796:Quinacrine / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*
chemical_list: D000098:Acetophenones; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D008555:Melitten; D003994:Bucladesine; D009637:Masoprocol; D011493:Protein Kinase C; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D011796:Quinacrine; D015232:Dinoprostone; C006821:4-bromophenacyl bromide; D007654:Ketoconazole