title: Inhibition of protein kinase C by ether-linked lipids is not correlated with their antineoplastic activity on WEHI-3B and R6X-B15 cells.
abstract: To test the hypothesis that the action of antineoplastic ether-linked lipids in leukemic cells is associated with their ability to inhibit protein kinase C (PKC), we have compared the effects of two ether-linked lipids, 1-O-hexadecyl-2-O-methyl-sn-glycero-3-phosphocholine (ET16-OCH3-GPC) and 1-O-hexadecyl-2-O-methyl-sn-glycero-3-(S-beta-D-1'- thioglucopyranosyl)-sn-glycerol (ET16-OCH3-beta-thio-Glc), on two different leukemic cell lines (WEHI-3B and R6X-B15). ET16-OCH3-GPC killed WEHI-3B cells with an EC50 value of 2.5 microM, whereas it was far less effective against R6X-B15 cells (EC50 = 40 microM). In contrast, the beta anomer of ET16-OCH3-beta-thio-Glc did not kill either cell line at concentrations up to 40 microM. Both ET16-OCH3-GPC and ET16-OCH3-thio-Glc inhibited 12-O-tetradecanoylphorbol 12,13-dibutyrate (TPA)-induced PKC translocation in both WEHI-3B and R6X-B15 cells. When WEHI-3B cells were first exposed to TPA, and then to ET16-OCH3-GPC, no significant decrease in PKC activity in the particulate fraction was noticed. When, however, the cells were first exposed to ET16-OCH3-GPC and then to TPA, the enzyme activity in the particulate fraction was decreased by 20-30%. A phorbol dibutyrate binding assay showed that the decrease in membrane-associated PKC activity and the increase in cytosolic PKC activity did not result from impeded enzyme translocation. These results suggest that the similar PKC inhibitory potency of ET16-OCH3-GPC and ET16-OCH3-beta-thio-Glc: (a) is not correlated with the widely different cytotoxicities of these agents and (b) is probably due to interference with the binding of diacylglycerol/phosphatidylserine or TPA to PKC. Taken together, these results suggest that the ether-linked lipids compete with diacylglycerol/phosphatidylserine or TPA for binding sites on PKC required for enzyme activation.
authors.affiliation: Department of Medicine, University of British Columbia, Vancouver, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Salari
authors.forename: P
authors.initials: P
authors.lastname: Dryden
authors.forename: R
authors.initials: R
authors.lastname: Davenport
authors.forename: S
authors.initials: S
authors.lastname: Howard
authors.forename: K
authors.initials: K
authors.lastname: Jones
authors.forename: R
authors.initials: R
authors.lastname: Bittman
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D002451:Cell Compartmentation; D002455:Cell Division / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D066298:In Vitro Techniques; D015240:Phorbol 12,13-Dibutyrate / Q000378:metabolism; D010742:Phospholipid Ethers / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D010742:Phospholipid Ethers; C026659:edelfosine; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate