MRP1 and glucosylceramide are coordinately over expressed and enriched in rafts during multidrug resistance acquisition in colon cancer cells.
Previously we have described a novel multidrug-resistant cell line, HT29(col), which displayed over expression of the multidrug-resistance protein 1 (MRP1) and an altered sphingolipid composition, including enhanced levels of glucosylceramide (GlcCer; Kok JW, Veldman RJ, Klappe K, Koning H, Filipeanu C, Muller M. Int J Cancer 2000;87:172-8). In our study, long-term screening revealed that, during colchicine-induced acquisition of multidrug resistance in a new HT29(col) cell line, increases in GlcCer occurred concomitantly with upregulation of MRP1 expression. Both MRP1 and GlcCer were found enriched in Lubrol-insoluble membrane domains. The expression of MRP1 and GlcCer were tightly correlated, as indicated also by a reversal of both at the later stage of colchicine consolidation. Resistance to colchicine was determined by MRP1, while glucosylceramide synthase (GCS) did not contribute: 1). Resistance was fully inhibited by MK571. 2). GCS expression and activity were not upregulated in HT29(col) cells. 3). Inhibition of GCS did not affect MRP1-mediated efflux function or sensitivity to colchicine. Instead, overall sphingolipid metabolism was upregulated through an increased rate of ceramide biosynthesis. In conclusion, upregulation of MRP1 occurs in concert with upregulation of GlcCer during multidrug-resistance acquisition, and both are enriched in rafts. The increased GlcCer pool does not directly modulate MRP1 function and cell survival.
Klappe
Karin
K
Department of Membrane Cell Biology, Groningen University Institute for Drug Exploration (GUIDE), University of Groningen, Groningen, The Netherlands.
Hinrichs
John W J
JW
Kroesen
Bart-Jan
BJ
Sietsma
Hannie
H
Kok
Jan Willem
JW
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000032:analysis / Q000502:physiology*; D017209:Apoptosis; D003078:Colchicine / Q000494:pharmacology; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D019008:Drug Resistance, Neoplasm; D005963:Glucosylceramides / Q000032:analysis / Q000502:physiology*; D005964:Glucosyltransferases / Q000037:antagonists & inhibitors / Q000502:physiology; D019073:HT29 Cells; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry*; D013107:Sphingolipids / Q000378:metabolism
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D005963:Glucosylceramides; D013107:Sphingolipids; D005964:Glucosyltransferases; C023246:ceramide glucosyltransferase; D003078:Colchicine