1-O-Octadecyl-2-O-methylglycerophosphocholine inhibits protein kinase C-dependent phosphorylation of endogenous proteins in MCF-7 cells.
Studies with leukaemic cells, based primarily on in vitro assays, have suggested that antitumour ether lipids have only a moderate effect on protein kinase C (PKC) activity, and, furthermore, inhibition of PKC is unlikely to be involved in the mechanism of inhibition of cell proliferation by these compounds. To determine if this is also the case for epithelial cancer cells, we examined the effect of 1-O-octadecyl-2-O-methylglycerophosphocholine (ET18-OCH3) on PKC-induced phosphorylation of endogenous proteins in MCF-7 cells under incubation conditions where the drug inhibited cell proliferation. As expected, stimulation of quiescent 32P-labelled MCF-7 cells with 1 microM PMA resulted in the phosphorylation of a number of proteins. The PMA-induced phosphorylation of the proteins was abolished by preincubation of the cells with Ro 31-8220 (5 microM) for 20 min, or 10 microg/ml ET18-OCH3 for 3 h before stimulation with PMA. Thus under incubation conditions where ET18-OCH3 inhibited the proliferation of MCF-7 cells, the ether lipid potently inhibited the activity of PKC in intact cells. This inhibition was unlikely to be due to the effect of the compound on PKC translocation since there was little effect of ET18-OCH3 on the translocation of the alpha, gamma and epsilon species of PKC. These results suggest that a role for the inhibition of PKC activity by ET18-OCH3 in the mechanism of inhibition of cell proliferation by ET18-OCH3 cannot yet be discounted in epithelial cancer cells. In addition, we also observed that ET18-OCH3 enhanced the phosphorylation of selected proteins under basal unstimulated conditions. Although some of these proteins were also observed to be phosphorylated in response to PMA stimulation, the phosphorylation induced by ET18-OCH3 was not inhibited by Ro 31-8220, indicating that this was not mediated by PKC.
Zhou
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Department of Biochemistry and Molecular Biology, University of Manitoba, 770 Bannatyne Avenue, Winnipeg R3E 0W3, Canada.
Arthur
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D001692:Biological Transport; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000494:pharmacology*; D010742:Phospholipid Ethers; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D004791:Enzyme Inhibitors; D009363:Neoplasm Proteins; D010713:Phosphatidylcholines; D010742:Phospholipid Ethers; C026659:edelfosine; D011493:Protein Kinase C