Influence of ceramide metabolism on P-glycoprotein function in immature acute myeloid leukemia KG1a cells.
Previous studies have emphasized the role of glucosylceramide (Glu-Cer) synthase in multidrug resistance (MDR) regulation. However, the mechanism by which the inhibition of this enzyme results in increased drug retention and cytotoxicity remains unclear. In this study, we investigated the respective role of ceramide (Cer) accumulation and Glu-Cer derivatives depletion in MDR reversal effect of 1-phenyl-2-decanoylamino-3-morpholino-1-propanolol (PDMP), a Glu-Cer synthase inhibitor. We show here that treatment with PDMP resulted in increased rhodamine 123 (Rh123) retention and potent chemosensitization of P-glycoprotein (P-gp)-expressing cells, including KG1a cells, KG1a/200 cells, K562/138 cells, and K562/mdr-1 cells. Metabolic studies revealed that PDMP induced not only time-dependent Cer accumulation but also reduction of all glycosylated forms of Cer, including Glu-Cer, lactosylceramide (Lac-Cer), monosialo ganglioside (GM3) and disialo ganglioside (GD3). The influence of these metabolites on P-gp function was investigated by measuring Rh123 retention in PDMP-treated cells. P-gp function was found to be stimulated only by the addition of gangliosides in all resistant cell lines, whereas Glu-Cer, Lac-Cer, and Cer had no effect. Moreover, in KG1a/200 cells, GD3 and, to a lesser extent, GM3 were found to phosphorylate P-gp on serine residues. Altogether, these results suggest that, at least in leukemic cells, gangliosides depletion accounts for PDMP-mediated MDR reversal effect, and that gangliosides are important P-gp regulators perhaps through their capacity to modulate P-gp phosphorylation.
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Isabelle
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Institut National de la Santé et de la Recherche Médicale (INSERM) E9910, Institut Claudius Régaud, Toulouse, France.
Lehne
Gustav
G
Beckstrøm
Karen Johanne
KJ
Maestre
Nicolas
N
Bettaïeb
Ali
A
Laurent
Guy
G
Lautier
Dominique
D
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000187:drug effects / Q000502:physiology*; D000208:Acute Disease; D015703:Antigens, CD; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002518:Ceramides / Q000494:pharmacology*; D003630:Daunorubicin / Q000494:pharmacology; D004347:Drug Interactions; D005732:Gangliosides / Q000494:pharmacology; D005963:Glucosylceramides / Q000494:pharmacology; D006801:Humans; D007790:Lactosylceramides / Q000494:pharmacology; D007951:Leukemia, Myeloid; D009025:Morpholines / Q000494:pharmacology*; D010766:Phosphorylation; D013110:Sphingosine / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014750:Vincristine / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D015703:Antigens, CD; D000970:Antineoplastic Agents; D002518:Ceramides; D005732:Gangliosides; D005963:Glucosylceramides; D007790:Lactosylceramides; D009025:Morpholines; C101954:N-caproylsphingosine; C009744:CDw17 antigen; D014750:Vincristine; C033110:RV 538; D013110:Sphingosine; D003630:Daunorubicin