QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services 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 Chen, Y. Articles by Porter, C. W. Search for Related Content PubMed PubMed Citation Articles by Chen, Y. Articles by Porter, C. W.

Small Interfering RNA Suppression of Polyamine Analog-Induced Spermidine/Spermine N¹-Acetyltransferase

Grace Cancer Drug Center, Roswell Park Cancer Institute, Buffalo, New York

N¹,N¹¹-diethylnorspermine (DENSPM) is a polyamine analog that down-regulates polyamine biosynthesis and potently upregulates the polyamine catabolic enzyme spermidine/spermine N¹-acetyltransferase (SSAT). In certain cells, such as SKMEL-28 human melanoma cells, induction of SSAT is associated with rapid apoptosis. In this study, we used small interfering RNA (siRNA) to examine the role of SSAT induction in mediating polyamine pool depletion and apoptosis. siRNA duplexes were designed to target three independent sites in the SSAT mRNA coding region (siSSAT). When transfected under nontoxic conditions, two of the duplexes selectively reduced basal SSAT mRNA in HEK-293 cells by >80% and prevented DENSPM-induced SSAT mRNA by 95% in SK-MEL-28 cells. Treatment of SK-MEL-28 cells with 10 µM DENSPM in the presence of 83 nM siSSAT selectively prevented the 1400-fold induction of SSAT activity by 90% and, in turn, prevented analog depletion of spermine (Spm) pools by 35%. siSSAT also prevented DENSPM-induced cytochrome c release and caspase-3 cleavage at 36 h and apoptosis at 48 h as measured by annexin V staining. Overall, the data directly link analog induction of SSAT to Spm pool depletion and to caspase-dependent apoptosis in DENSPM-treated SK-MEL-28 cells. This represents the first use of siRNA technology directed toward a polyamine gene and the first unequivocal demonstration that SSAT induction initiates events leading to polyamine analog-induced apoptosis.

Address correspondence to: Dr. Carl W. Porter, Grace Cancer Drug Center, Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263. E-mail: carl.porter{at}roswellpark.org

This article has been cited by other articles:

				D. L. Kramer, P. Diegelman, J. Jell, S. Vujcic, S. Merali, and C. W. Porter Polyamine Acetylation Modulates Polyamine Metabolic Flux, a Prelude to Broader Metabolic Consequences J. Biol. Chem., February 15, 2008; 283(7): 4241 - 4251. [Abstract] [Full Text] [PDF]

				W. L. Allen, E. G. McLean, J. Boyer, A. McCulla, P. M. Wilson, V. Coyle, D. B. Longley, R. A. Casero Jr., and P. G. Johnston The role of spermidine/spermine N1-acetyltransferase in determining response to chemotherapeutic agents in colorectal cancer cells Mol. Cancer Ther., January 1, 2007; 6(1): 128 - 137. [Abstract] [Full Text] [PDF]

				A. Pledgie, Y. Huang, A. Hacker, Z. Zhang, P. M. Woster, N. E. Davidson, and R. A. Casero Jr. Spermine Oxidase SMO(PAOh1), Not N1-Acetylpolyamine Oxidase PAO, Is the Primary Source of Cytotoxic H2O2 in Polyamine Analogue-treated Human Breast Cancer Cell Lines J. Biol. Chem., December 2, 2005; 280(48): 39843 - 39851. [Abstract] [Full Text] [PDF]

				J. M. Tucker, J. T. Murphy, N. Kisiel, P. Diegelman, K. W. Barbour, C. Davis, M. Medda, L. Alhonen, J. Janne, D. L. Kramer, et al. Potent Modulation of Intestinal Tumorigenesis in Apcmin/+ Mice by the Polyamine Catabolic Enzyme Spermidine/Spermine N1-acetyltransferase Cancer Res., June 15, 2005; 65(12): 5390 - 5398. [Abstract] [Full Text] [PDF]

				N. Martinez, M. Sanchez-Beato, A. Carnero, V. Moneo, J. C. Tercero, I. Fernandez, M. Navarrete, J. Jimeno, and M. A. Piris Transcriptional signature of Ecteinascidin 743 (Yondelis, Trabectedin) in human sarcoma cells explanted from chemo-naive patients Mol. Cancer Ther., May 1, 2005; 4(5): 814 - 823. [Abstract] [Full Text] [PDF]

				K. Kee, B. A. Foster, S. Merali, D. L. Kramer, M. L. Hensen, P. Diegelman, N. Kisiel, S. Vujcic, R. V. Mazurchuk, and C. W. Porter Activated Polyamine Catabolism Depletes Acetyl-CoA Pools and Suppresses Prostate Tumor Growth in TRAMP Mice J. Biol. Chem., September 17, 2004; 279(38): 40076 - 40083. [Abstract] [Full Text] [PDF]

				R. Chaturvedi, Y. Cheng, M. Asim, F. I. Bussiere, H. Xu, A. P. Gobert, A. Hacker, R. A. Casero Jr., and K. T. Wilson Induction of Polyamine Oxidase 1 by Helicobacter pylori Causes Macrophage Apoptosis by Hydrogen Peroxide Release and Mitochondrial Membrane Depolarization J. Biol. Chem., September 17, 2004; 279(38): 40161 - 40173. [Abstract] [Full Text] [PDF]

				S. Hector, C. W. Porter, D. L. Kramer, K. Clark, J. Prey, N. Kisiel, P. Diegelman, Y. Chen, and L. Pendyala Polyamine catabolism in platinum drug action: Interactions between oxaliplatin and the polyamine analogue N1,N11-diethylnorspermine at the level of spermidine/spermine N1-acetyltransferase Mol. Cancer Ther., July 1, 2004; 3(7): 813 - 822. [Abstract] [Full Text] [PDF]

				K. Kee, S. Vujcic, S. Merali, P. Diegelman, N. Kisiel, C. T. Powell, D. L. Kramer, and C. W. Porter Metabolic and Antiproliferative Consequences of Activated Polyamine Catabolism in LNCaP Prostate Carcinoma Cells J. Biol. Chem., June 25, 2004; 279(26): 27050 - 27058. [Abstract] [Full Text] [PDF]