Cytotoxicity and metabolism of alkyl phospholipid analogues in neoplastic cells.
The cytotoxic response of several types of neoplastic cells to analogues of unnatural alkyl phospholipids (e.g., rac-1-hexadecyl-2-methoxy-glycero-3-phosphocholine) has been partially attributed to their accumulation as a result of the low activity of the alkyl cleavage enzyme (a tetrahydropteridine-dependent monooxygenase) in tumor cells. We tested this possibility by comparing the alkyl cleavage enzyme activity in cells that exhibit differences in sensitivity toward the cytotoxic effects of the rac-1-hexadecyl-2-methoxy-glycero-3-phosphocholine. Human promyelocytic leukemia cells (HL-60), a cell line highly sensitive to the cytotoxic alkyl phospholipid analogue, possessed an alkyl cleavage enzyme activity (0.25 pmol/min/microgram protein) similar to that found in three cell types known to be relatively resistant to the cytotoxic activity of the analogue: immature human promyeloblastic leukemia cells (K562) (0.22 pmol/min/microgram protein), human polymorphonuclear neutrophils (0.34 pmol/min/microgram protein), and Madin-Darby canine kidney cells (0.37 pmol/min/microgram protein). Moreover, our results indicate that the cytotoxic rac-1-octadecyl-2-methoxy-glycero-3-phosphocholine analogue is not a substrate for the alkyl cleavage enzyme with an active microsomal preparation of the enzyme from rat liver; cleavage of this analogue was 200-fold less than the rate obtained with 1-octadecylglycerol as substrate. In cultures of either sensitive or resistant type cells, approximately 90% of the added rac-1-[9',10'-3H]octadecyl-2-methoxy-glycero-3-phosphocholine was not metabolized during a 24-h incubation. The amount of radiolabel in fatty acids, a major product of alkyl cleavage activity, was small, and essentially identical amounts were produced in all four cell types [3.1 +/- 0.2% (SD)]. These data indicate that differences in the cellular activities of the alkyl cleavage enzyme are not responsible for the differential cytotoxic responses between normal and specific types of neoplastic cells toward rac-1-octadecyl-2-methoxy-glycero-3-phosphocholine. On the other hand, the cellular uptake of the alkyl phospholipids could be a factor in explaining the cytotoxic response of certain tumor cells, since more radiolabeled 1-octadecyl-2-methoxy-glycero-3-phosphocholine was associated with the susceptible HL-60 cells than with the resistant cell types. Autoradiography revealed that the radiolabeled 2-methoxy analogue accumulates at the periphery of HL-60 leukemia cells, whereas the label was more uniformly distributed in polymorphonuclear neutrophils and K562 cells.(ABSTRACT TRUNCATED AT 400 WORDS)
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism* / Q000633:toxicity; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004285:Dogs; D006801:Humans; D008244:Lysophosphatidylcholines / Q000378:metabolism* / Q000633:toxicity; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D010742:Phospholipid Ethers; D010972:Platelet Activating Factor / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity
D000970:Antineoplastic Agents; D008244:Lysophosphatidylcholines; D010742:Phospholipid Ethers; D010972:Platelet Activating Factor; C026659:edelfosine; C050186:1-hexadecyl-2-methoxyglycero-3-phosphocholine; D006899:Mixed Function Oxygenases; C020678:glyceryl-ether monooxygenase