Premium
Differential Effects of Glycolipid Biosynthesis Inhibitors on Ceramide‐Induced Cell Death in Neuroblastoma Cells
Author(s) -
Bieberich Erhard,
Freischütz Bettina,
Suzuki Minoru,
Yu Robert K
Publication year - 1999
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1999.0721040.x
Subject(s) - ceramide , glycolipid , ceramide synthase , neuroblastoma , biochemistry , apoptosis , sphingolipid , programmed cell death , intracellular , biology , cell culture , chemistry , microbiology and biotechnology , genetics
: An in vitro model of Gaucher's disease in murineneuroblastoma X rat glioma NG108‐15 cells was used to investigate thephysiological effects of two specific inhibitors of glucosylceramide synthase, d,l‐threo ‐1‐phenyl‐2‐decanoylamino‐3‐morpholino‐1‐propanol( d,l ‐PDMP) and N ‐butyldeoxynojirimycin (NB‐DNJ), which havebeen suggested as agents for treatment of glycolipid storage disorders.Incubation of NG108‐15 cells with conduritol‐B‐epoxide, a covalent inhibitorof glucosylce‐ramidase, raised the intracellular concentration ofglucosylceramide (GC) by more than fourfold, indicating a glycolipidcomposition equivalent to that of Gaucher's cells. The level of GC wasdecreased, and the cells were depleted of gangliosides by postincubation with d,l ‐PDMP or NB‐DNJ. Treatment with d,l ‐PDMP, but not withNB‐DNJ, resulted in a dose‐dependent reduction of the growth rate andeventually caused cell death in NG108‐15 cells on reaching confluency. An insitu detection assay using terminal nucleotidyltransferase indicated that celldegeneration was accompanied by apoptosis. Lipid analysis by high‐performanceTLC revealed that on incubation with d,l ‐PDMP, but not with NB‐DNJ,the concentration of endogenous ceramide was elevated by threefold. Ceramideelevation and apoptosis were also observed when NG108‐15 cells were incubatedwith daunorubicin, which was previously reported to induce programmed celldeath by stimulation of ceramide synthesis. Structural characterization byHPLC and subsequent laser desorption mass spectrometry revealed that theendogenous ceramide contained fatty acids with chain lengths ranging from C14: 0 to C24 : 0. The results indicate that elevation of levels of theseceramide species by incubation with d,l ‐PDMP or daunorubicin inducesprogrammed cell death in NG108‐15 cells. Because ceramide accumulation andcell death were not observed on incubation with NB‐DNJ, its use is suggestedto be less toxic than that of d,l ‐PDMP for treatment of Gaucher's disease and other sphingolipid storage disorders.