Directed shift of fatty acids from phospholipids to triacylglycerols in HL-60 cells induced by nanomolar concentrations of triethyl lead chloride: involvement of a pertussis toxin-sensitive pathway.
Triethyl lead chloride (Et3PbCl) was found to induce a shift of fatty acids from membrane phospholipids to triacylglycerols in the human promyelocytic leukemia cell line HL-60. High concentrations of Et3PbCl (greater than 10 microM) caused a substantial liberation of [14C]arachidonic acid within 10 to 20 min in dimethyl sulfoxide-differentiated cells, comparable to the effect of the calcium ionophore A23187 (10 microM). Following liberation of arachidonic acid, its metabolites could be detected. Prolongation of the incubation time and reduction of Et3PbCl concentration resulted in a shift of fatty acids from phospholipids to triacylglycerols. Deacylation of phospholipids and reacylation into phospholipids and triacylglycerols were in equilibrium when the cells were treated with Et3PbCl at concentrations of less than or equal to 10 microM for 5 hr or less than or equal to 1 microM for 24 hr; no increase of free fatty acids could be observed, and the loss of fatty acids within the phospholipids was equivalent to the increase of fatty acid content within the triacylglycerols. Moreover, under these conditions, no loss of viability was seen after 24 hr, as compared with untreated differentiated cells. This concentration- and time-dependent effect of Et3PbCl might be due to a stimulated liberation of fatty acids via phospholipase A2, because this stimulation could be totally prevented by the phospholipase inhibitors quinacrine and p-bromophenacylbromide. Additionally, pretreatment of differentiated HL-60 cells with pertussis toxin resulted in a drastic reduction of [14C]arachidonic acid liberation when cells were stimulated with Et3PbCl. These results suggest the involvement of a pertussis toxin-sensitive GTP-binding protein and of a signal transduction mechanism during stimulated fatty acid release; release does not seem to be via a direct stimulation of phospholipase activity by the lead compound.
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Kernforschungszentrum Karlsruhe, Institut für Genetik und für Toxikologie, Federal Republic of Germany.
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D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000037:antagonists & inhibitors; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D004789:Enzyme Activation; D005227:Fatty Acids / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D006801:Humans; D007942:Leukemia, Experimental / Q000378:metabolism / Q000473:pathology; D009942:Organometallic Compounds / Q000633:toxicity*; D037342:Pertussis Toxin; D010740:Phospholipases / Q000037:antagonists & inhibitors; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism*; D013849:Thimerosal / Q000494:pharmacology; D014280:Triglycerides / Q000378:metabolism*; D014407:Tumor Cells, Cultured / Q000378:metabolism*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D001095:Arachidonic Acids; D005227:Fatty Acids; D009942:Organometallic Compounds; D010743:Phospholipids; D014280:Triglycerides; D010566:Virulence Factors, Bordetella; D013849:Thimerosal; D016718:Arachidonic Acid; C010360:triethyllead; D008243:1-Acylglycerophosphocholine O-Acyltransferase; D037342:Pertussis Toxin; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D019204:GTP-Binding Proteins