title: Relationship between arachidonate--phospholipid remodeling and apoptosis.
abstract: Our previous studies reveal that three structurally distinct inhibitors of the enzyme CoA-independent transacylase, including the antiproliferative alkyllysophospholipid ET-18-O-CH3, induce programmed cell death (apoptosis) in the promyelocytic cell line HL-60. The objective of the current study was to better elucidate the mechanism responsible for apoptosis. CoA-IT is an enzyme believed to be responsible for the remodeling of long chain polyunsaturated fatty acids like arachidonate between the phospholipids of mammalian cells. The chronic (24-48 h) treatment of HL-60 cells with all three CoA-IT inhibitors resulted in the inhibition of the remodeling of labeled arachidonate from choline- into ethanolamine-containing phospholipid molecular species. GC-MS analysis of the fatty acids in phospholipids revealed that CoA-IT inhibitor treatment induced a marked loss of arachidonate-containing phosphatidylethanolamine and an increase in arachidonate-containing phosphatidylcholine. This redistribution was specific to arachidonate since the mass distribution of linoleic acid in glycerolipids was not affected. In spite of the dramatic redistribution of arachidonate, the total cellular arachidonate content was not altered nor was the relative distribution of total phospholipid classes. The increase of arachidonate in phosphatidylcholine was specifically due to an increase in 1-acyl-2-arachidonoyl-sn-glycero-3-phosphocholine species, whereas the loss of arachidonate in PE was from both 1-acyl- and 1-alk-1-enyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine species. The incubation of cells with exogenous arachidonic acid or ethanolamine did not reverse the inhibition of proliferation induced by CoA-IT inhibitor treatment. Incubation with CoA-IT inhibitors also induced the characteristic cytoplasmic and nuclear changes associated with apoptosis as assessed by transmission electron microscopy and DNA fragmentation as determined by flow cytometry. Taken together, these data show that apoptosis in HL-60 cells, induced by blocking arachidonate-phospholipid remodeling, is correlated with a redistribution of arachidonate in membrane phospholipids and suggest that such alterations represent a signal which controls the capacity of cells to proliferate.
authors.affiliation: Section on Pulmonary and Critical Care Medicine, Bowman Gray School of Medicine, Winston-Salem, North Carolina 27157-1054, USA.
authors.forename: M E
authors.initials: ME
authors.lastname: Surette
authors.forename: J D
authors.initials: JD
authors.lastname: Winkler
authors.forename: A N
authors.initials: AN
authors.lastname: Fonteh
authors.forename: F H
authors.initials: FH
authors.lastname: Chilton
mesh_terms: D000217:Acyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017209:Apoptosis / Q000502:physiology*; D016718:Arachidonic Acid / Q000378:metabolism*; D001557:Benzenesulfonates / Q000494:pharmacology; D002452:Cell Count / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D019787:Linoleic Acid; D008041:Linoleic Acids / Q000032:analysis / Q000378:metabolism; D008854:Microscopy, Electron; D009943:Organophosphorus Compounds / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D010742:Phospholipid Ethers / Q000494:pharmacology; D010743:Phospholipids / Q000378:metabolism*; D014508:Urea / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: C093344:2-(2-(3-(4-chloro-3-(trifluoromethyl)phenyl)ureido)-4-(trifluoromethyl)phenoxy)-4,5-dichlorobenzenesulfonic acid; D001557:Benzenesulfonates; D004791:Enzyme Inhibitors; D007093:Imidazoles; D008041:Linoleic Acids; D009943:Organophosphorus Compounds; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010742:Phospholipid Ethers; D010743:Phospholipids; C093303:SK&F 98625; C026659:edelfosine; D016718:Arachidonic Acid; D014508:Urea; D019787:Linoleic Acid; D000217:Acyltransferases; C039766:arachidonyl transacylase