This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wang, H. Articles by Cabot, M. C. Search for Related Content PubMed PubMed Citation Articles by Wang, H. Articles by Cabot, M. C.

N-(4-Hydroxyphenyl)retinamide Elevates Ceramide in Neuroblastoma Cell Lines by Coordinate Activation of Serine Palmitoyltransferase and Ceramide Synthase¹

John Wayne Cancer Institute at Saint John’s Health Center, Santa Monica, California 90404 [H. W., M. C. C.]; Division of Hematology-Oncology, Children’s Hospital, Los Angeles, California 90027 [B. J. M., C. P. R.]; and Departments of Pediatrics [B. J. M., C. P. R.], Cell and Neurobiology [B. J. M.], and Pathology [C. P. R.], University of Southern California Keck School of Medicine, Los Angeles, California 90033

The retinoid N-(4-hydroxyphenyl)retinamide (4-HPR; fenretinide) is cytotoxic to a variety of cancer cell lines, and we previously showed an association between ceramide generation and 4-HPR cytotoxicity for neuroblastoma cell lines (B. J. Maurer et al., J. Natl. Cancer Inst. (Bethesda), 91: 1138–1146, 1999). Here we determine whether the increased ceramide mediated by 4-HPR in the CHLA-90 human neuroblastoma cell line results from de novo ceramide synthesis. Treatment of CHLA-90 with 4-HPR for 2 h, in the presence of [³H]palmitic acid, caused sequential formation of [³H]sphinganine (220% over control) and [³H]ceramide (160% over control), with sphinganine returning to baseline at 4 h, and ceramide continuing to increase (215% over control). 4-HPR treatment did not accelerate cellular decay of sphingomyelin. Preincubation of cells with either L-cycloserine, an inhibitor of serine palmitoyltransferase (SPT), or fumonisin B₁, an inhibitor of ceramide synthase, retarded ceramide formation in response to 4-HPR treatment, although sphinganine was still generated when 4-HPR and FB₁ were present. Data from in vitro enzyme assays using microsomes showed that preexposure of intact cells to 4-HPR resulted in a time (175% over control; 6 h)- and dose-dependent increase (173% over control; 10 µM) in SPT activity as well as a time (265% over control)- and dose-dependent increase (215% above control; 10 µM) in ceramide synthase activity. Our results show that 4-HPR-mediated ceramide generation is derived from the de novo synthetic pathway by coordinate activation of SPT and ceramide synthase. Knowledge of these biochemical events is of utility when downstream modulators of ceramide metabolism are used to heighten the cytotoxic response to chemotherapy.

This article has been cited by other articles:

				S. Schiffmann, J. Sandner, R. Schmidt, K. Birod, I. Wobst, H. Schmidt, C. Angioni, G. Geisslinger, and S. Grosch The selective COX-2 inhibitor celecoxib modulates sphingolipid synthesis J. Lipid Res., January 1, 2009; 50(1): 32 - 40. [Abstract] [Full Text] [PDF]

				D. E. Saslowsky, N. Tanaka, K. P. Reddy, and W. I. Lencer Ceramide activates JNK to inhibit a cAMP-gated K+ conductance and Cl- secretion in intestinal epithelia FASEB J, January 1, 2009; 23(1): 259 - 270. [Abstract] [Full Text] [PDF]

				H. Wang, B. J. Maurer, Y.-Y. Liu, E. Wang, J. C. Allegood, S. Kelly, H. Symolon, Y. Liu, A. H. Merrill, Jr., V. Gouaze-Andersson, et al. N-(4-Hydroxyphenyl)retinamide increases dihydroceramide and synergizes with dimethylsphingosine to enhance cancer cell killing Mol. Cancer Ther., September 1, 2008; 7(9): 2967 - 2976. [Abstract] [Full Text] [PDF]

				S. T. Pruett, A. Bushnev, K. Hagedorn, M. Adiga, C. A. Haynes, M. C. Sullards, D. C. Liotta, and A. H. Merrill Jr. Thematic Review Series: Sphingolipids. Biodiversity of sphingoid bases ("sphingosines") and related amino alcohols J. Lipid Res., August 1, 2008; 49(8): 1621 - 1639. [Abstract] [Full Text] [PDF]

				P. G. Sreekumar, J. Zhou, J. Sohn, C. Spee, S. J. Ryan, B. J. Maurer, R. Kannan, and D. R. Hinton N-(4-hydroxyphenyl) Retinamide Augments Laser-Induced Choroidal Neovascularization in Mice Invest. Ophthalmol. Vis. Sci., March 1, 2008; 49(3): 1210 - 1220. [Abstract] [Full Text] [PDF]

				J. M. Kraveka, L. Li, Z. M. Szulc, J. Bielawski, B. Ogretmen, Y. A. Hannun, L. M. Obeid, and A. Bielawska Involvement of Dihydroceramide Desaturase in Cell Cycle Progression in Human Neuroblastoma Cells J. Biol. Chem., June 8, 2007; 282(23): 16718 - 16728. [Abstract] [Full Text] [PDF]

				M. G. Villani, V. Appierto, E. Cavadini, A. Bettiga, A. Prinetti, M. Clagett-Dame, R. W. Curley, and F. Formelli 4-Oxo-Fenretinide, a Recently Identified Fenretinide Metabolite, Induces Marked G2-M Cell Cycle Arrest and Apoptosis in Fenretinide-Sensitive and Fenretinide-Resistant Cell Lines. Cancer Res., March 15, 2006; 66(6): 3238 - 3247. [Abstract] [Full Text] [PDF]

				A. Schulz, T. Mousallem, M. Venkataramani, D.-A. Persaud-Sawin, A. Zucker, C. Luberto, A. Bielawska, J. Bielawski, J. C. M. Holthuis, S. M. Jazwinski, et al. The CLN9 Protein, a Regulator of Dihydroceramide Synthase J. Biol. Chem., February 3, 2006; 281(5): 2784 - 2794. [Abstract] [Full Text] [PDF]

				E. Y. Park, A. Dillard, E. A. Williams, E. T. Wilder, M. R. Pepper, and M. A. Lane Retinol Inhibits the Growth of All-Trans-Retinoic Acid-Sensitive and All-Trans-Retinoic Acid-Resistant Colon Cancer Cells through a Retinoic Acid Receptor-Independent Mechanism Cancer Res., November 1, 2005; 65(21): 9923 - 9933. [Abstract] [Full Text] [PDF]

				E. A. Sausville Indifferently Pursued or Unowned Drugs: Who Should Lead Where Companies Do Not Tread? J. Clin. Oncol., March 20, 2005; 23(9): 1796 - 1798. [Full Text] [PDF]

				C. M. Finnegan, S. S. Rawat, A. Puri, J. M. Wang, F. W. Ruscetti, and R. Blumenthal Ceramide, a target for antiretroviral therapy PNAS, October 26, 2004; 101(43): 15452 - 15457. [Abstract] [Full Text] [PDF]

				P. E. Lovat, F. Di Sano, M. Corazzari, B. Fazi, R. P. Donnorso, A. D. J. Pearson, A. G. Hall, C. P. F. Redfern, and M. Piacentini Gangliosides Link the Acidic Sphingomyelinase-Mediated Induction of Ceramide to 12-Lipoxygenase-Dependent Apoptosis of Neuroblastoma in Response to Fenretinide J Natl Cancer Inst, September 1, 2004; 96(17): 1288 - 1299. [Abstract] [Full Text] [PDF]

				A. Prinetti, L. Basso, V. Appierto, M. G. Villani, M. Valsecchi, N. Loberto, S. Prioni, V. Chigorno, E. Cavadini, F. Formelli, et al. Altered Sphingolipid Metabolism in N-(4-Hydroxyphenyl)- retinamide-resistant A2780 Human Ovarian Carcinoma Cells J. Biol. Chem., February 14, 2003; 278(8): 5574 - 5583. [Abstract] [Full Text] [PDF]

				A. Erdreich-Epstein, L. B. Tran, N. N. Bowman, H. Wang, M. C. Cabot, D. L. Durden, J. Vlckova, C. P. Reynolds, M. F. Stins, S. Groshen, et al. Ceramide Signaling in Fenretinide-induced Endothelial Cell Apoptosis J. Biol. Chem., December 13, 2002; 277(51): 49531 - 49537. [Abstract] [Full Text] [PDF]

				G. M. Jenkins, L. A. Cowart, P. Signorelli, B. J. Pettus, C. E. Chalfant, and Y. A. Hannun Acute Activation of de Novo Sphingolipid Biosynthesis upon Heat Shock Causes an Accumulation of Ceramide and Subsequent Dephosphorylation of SR Proteins J. Biol. Chem., November 1, 2002; 277(45): 42572 - 42578. [Abstract] [Full Text] [PDF]

				H. Wang, A. E. Giuliano, and M. C. Cabot Enhanced de Novo Ceramide Generation through Activation of Serine Palmitoyltransferase by the P-Glycoprotein Antagonist SDZ PSC 833 in Breast Cancer Cells Mol. Cancer Ther., July 1, 2002; 1(9): 719 - 726. [Abstract] [Full Text] [PDF]