Ceramide Reduction and Transcriptional Up-Regulation of Glucosylceramide Synthase through Doxorubicin-Activated Sp1 in Drug-Resistant HL-60/ADR Cells
Open Access
- 1 September 2004
- journal article
- Published by American Association for Cancer Research (AACR) in Cancer Research
- Vol. 64 (17), 6271-6279
- https://doi.org/10.1158/0008-5472.can-03-1476
Abstract
Treatment with doxorubicin (DOX) induced apoptosis with an increase of ceramide content in drug-sensitive HL-60 cells, but not in drug-resistant HL-60/ADR cells. In HL-60/ADR cells (but not in HL-60 cells), the levels of mRNA, protein, and activity in glucosylceramide synthase (GCS), which converts ceramide to glucosylceramide, were up-regulated in response to DOX. Thus, abrogation of apoptosis in HL-60/ADR cells might be involved in ceramide reduction through DOX-induced up-regulation of GCS function. Because we reported that a GC-rich/Sp1 promoter binding region was of importance in the regulation of GCS expression, the role of Sp1 in DOX-induced up-regulation of GCS and apoptosis was investigated. DOX induced Sp1 activation in HL-60/ADR cells, as assessed by Sp1 gel shift and promoter-luciferase reporter assays, whereas transfection of double-stranded oligodeoxynucleotides (ODNs) containing a GC-rich/Sp1 region (Sp1 decoy ODNs) inhibited DOX-induced Sp1 activation. In addition, DOX-increased mRNA and enzyme activity in GCS were inhibited by Sp1 decoy, in conjunction with corresponding elevations of ceramide content. Moreover, DOX-induced apoptotic cell death was significantly increased in Sp1 decoy ODN-transfected HL-60/ADR cells over mock-transfected HL-60/ADR cells. Together, the results suggest that transcriptional up-regulation of GCS through DOX-induced activation of Sp1 is one potential mechanism to regulate ceramide increase and apoptosis in HL-60/ADR cells.Keywords
This publication has 29 references indexed in Scilit:
- Common themes in the pathogenesis of acute myeloid leukemiaOncogene, 2001
- Role of c-jun Expression Increased by Heat Shock- and Ceramide-activated Caspase-3 in HL-60 Cell ApoptosisPublished by Elsevier BV ,2000
- Glycosylation of Ceramide Potentiates Cellular Resistance to Tumor Necrosis Factor-α-Induced ApoptosisExperimental Cell Research, 1999
- Molecular Cloning and Characterization of the Mouse Ceramide Glucosyltransferase GeneBiochemical and Biophysical Research Communications, 1998
- Ceramide-induced Translocation of Protein Kinase C-δ and -ϵ to the CytosolJournal of Biological Chemistry, 1997
- Life, death, and the pursuit of apoptosis.Genes & Development, 1996
- Purification and Characterization of UDP-glucose:Ceramide Glucosyltransferase from Rat Liver Golgi MembranesPublished by Elsevier BV ,1996
- Ceramide synthase mediates daunorubicin-induced apoptosis: An alternative mechanism for generating death signalsCell, 1995
- Programmed Cell Death Induced by CeramideScience, 1993
- A RAPID METHOD OF TOTAL LIPID EXTRACTION AND PURIFICATIONCanadian Journal of Biochemistry and Physiology, 1959