Separation of several 5-fluorouracil metabolites in various melanoma cell lines. Evidence for the synthesis of 5-fluorouracil-nucleotide sugars.
5-Fluorouracil (5FU) metabolism was studied in intact cancer cells kept in suspension by incubation with tritiated 5FU. Metabolites were analyzed using various chromatographic procedures, including a one-directional separation on PEI-cellulose sheets, which separated 5FU from the mono-, di- and triphosphate forms and from nucleotide sugars. The monophosphate ester was present as FdUMP, as could be demonstrated with another chromatographic procedure. In the human melanoma cell lines IGR3 and M5 the main metabolite of 5FU was 5-fluorouridine, while in the murine B16 melanoma only a small amount of 5-fluorouridine was formed. In B16 cells more 5FU label was present as the triphosphate ester while in M5 cells more FdUMP was formed. With all three cell lines 5FU was incorporated into RNA; this incorporation was stimulated by 1 mM N-(phosphonacetyl)-L-aspartate (PALA). PALA did not significantly affect the conversion of 5FU into other metabolites, nor did it affect the incorporation of 5FU into DNA. A 5FU-nucleotide sugar, present as the diphosphate-glucose, was a predominant 5FU-metabolite in M5 cells but not in the other cell lines. Its identity was confirmed by thin-layer chromatography and high-performance liquid chromatography. Its possible function is discussed.
D000818:Animals; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D002845:Chromatography; D003856:Deoxyuracil Nucleotides / Q000096:biosynthesis; D005468:Fluorodeoxyuridylate / Q000096:biosynthesis; D005472:Fluorouracil / Q000378:metabolism*; D006801:Humans; D008545:Melanoma / Q000378:metabolism*; D051379:Mice; D010746:Phosphonoacetic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D012334:RNA, Neoplasm / Q000096:biosynthesis; D014500:Uracil Nucleotides / Q000096:biosynthesis*; D014529:Uridine / Q000031:analogs & derivatives / Q000096:biosynthesis; D014544:Uridine Triphosphate / Q000031:analogs & derivatives / Q000096:biosynthesis
D003856:Deoxyuracil Nucleotides; D012334:RNA, Neoplasm; D014500:Uracil Nucleotides; D005468:Fluorodeoxyuridylate; D001224:Aspartic Acid; C025635:5-fluorouridine 5'-triphosphate; C001943:5-fluorouridine; C013195:sparfosic acid; D010746:Phosphonoacetic Acid; D005472:Fluorouracil; D014544:Uridine Triphosphate; D014529:Uridine