Abstract
Purpose
Our previous studies showed that combined treatment of oxaliplatin and N 1 , N 11diethyl-norspermine (DENSPM) results in massive induction of spermidine/spermine N 1-acetyltransferase (SSAT) mRNA and activity. Since oxaliplatin and 5-fluorouracil (5FU) are used clinically in treatment of colorectal cancers, this study examines the effect of adding DENSPM to oxaliplatin/5FU combination on SSAT and spermine oxidase (SMO) in HCT-116 cells.
Methods
HCT-116 cells were treated with clinically relevant concentrations of drugs for 20 h followed by 24 h in drug free medium. SSAT and SMO mRNA and protein were assayed by QRT-PCR and Westerns respectively; polyamine pools were measured by HPLC. SSAT and SMO mRNA in tumor biopsies from patients with rectal cancer receiving oxaliplatin, capecitabine and radiation were measured by QRT-PCR.
Results
Oxaliplatin + 5FU + DENSPM produced significantly higher levels of SSAT and SMO mRNA, protein and activity than those seen with oxaliplatin+5FU with a significant depletion of cellular spermine and spermidine pools. Oxaliplatin/DENSPM was superior to 5FU/DENSPM in SSAT induction but similar for SMO. Oxaliplatin + DENSPM revealed synergistic growth inhibition at >IC50 concentrations and antagonism at <IC50. SMO and SSAT induction occurred in 60 and 30% of the patient samples examined.
Conclusions
These studies demonstrated that combining DENSPM with oxaliplatin + 5FU provides an added benefit by aiming at the clinically relevant therapeutic target, the polyamine catabolism. Further, we show for the first time, that SMO and SSAT induction could be measured in tumor biopsies in patients receiving chemo-radiation. Optimization of treatment conditions in vivo should facilitate a clinical evaluation of the three drug combination.
Similar content being viewed by others
Abbreviations
- DENSPM:
-
N 1 N 11-Diethylnorspermine
- ERCC-1:
-
Excision cross-complementing gene-1
- 5FU:
-
5-Fluorouracil
- dUMP:
-
Deoxyuridine monophosphate
- Pt:
-
Platinum
- Put:
-
Putrescine
- QRT-PCR:
-
Quantitative RT-PCR
- Spd:
-
Spermidine
- Spm:
-
Spermine
- SSAT:
-
Spermidine/spermine N 1-acetyltransferase (also known as SSAT-1)
- SMO:
-
Spermine oxidase
- SRB:
-
Sulforhodamine-B
- TMP:
-
Thymidine monophosphosphate
- TP:
-
Thymidine phosphorylase
- TS:
-
Thymidylate synthase
- FdUMP:
-
5-Fluorouridine monophosphate
References
Abeloff MD, Rosen ST, Luk GD, Baylin SB, Zeltzman M, Sjoerdsma A (1986) Phase II trials of a-difluoromethylornithine, an inhibitor of polyamine synthesis in advanced small cell lung cancer and colon cancer. Cancer Treat Rep 70:843–845
Alhonen L, Pietila M, Halmekyto M, Kramer DL, Janne J, Porter CW (1999) Transgenic mice with activated polyamine catabolism due to overexpression of spermidine/spermine N1-acetyltransferase show enhanced sensitivity to the polyamine analog, N1, N11-diethylnorspermine. Mol Pharmacol 55:693–698
Allen WL, McLean EG, Boyer J, McCulla A, Wilson PM, Coyle V, Longley DB, Casero RA Jr, Johnston PG (2007) The role of spermidine/spermine N1-acetyltransferase in determining response to chemotherapeutic agents in colorectal cancer cells. Mol Cancer Ther 6:128–137
Boyer J, Allen WL, McLean EG, Wilson PM, McCulla A, Moore S, Longley DB, Caldas C, Johnston PG (2006) Pharmacogenomic identification of novel determinants of response to chemotherapy in colon cancer. Cancer Res 66:2765–2777
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 52:5115–5118
Casero RA Jr, Wang Y, Stewart TM, Devereux W, Hacker A, Wang Y, Smith R, Woster PM (2003) The role of polyamine catabolism in anti-tumour drug response. Biochem Soc Trans 31:361–365
Chen Y, Kramer D, Jell J, Vujcic S, Porer CW (2003) siRNA suppression of polyamine analogue-induced spermidine/spermine N1-acetyltransferase. Mol Pharmacol 64:1153–1159
Chen Y, Kramer DL, Diegelman P, Vujcic S, Porter CW (2001) Apoptotic signaling in polyamine analogue-treated SK-MEL-28 human melanoma cells. Can Res 61:6437–6444
Choi W, Gerner EW, Ramdas L, Dupart J, Carew J, Proctor L, Huang P, Zhang W, Hamilton SR (2004) Combination of 5-fluorouracil and N1,N11-diethylnorspermine markedly activates spermidine/spermine N1-acetyltransferase expression, depletes polyamines, and synergistically induces apoptosis in colon carcinoma cells. J Biol Chem 280:3295–3304
Choi W, Proctor L, Xia Q, Feng Y, Gerner EW, Chiao PJ, Hamilton SR, Zhang W (2006) Inactivation of IkappaB contributes to transcriptional activation of spermidine/spermine N(1)-acetyltransferase. Mol Carcinog 45:685–693
Coleman CS, Pegg AE (2001) Polyamine analogues inhibit the ubiquitination of spermidine/spermine N1-acetyltransferase and prevent its targeting to the proteasome for degradation. Biochem J 358:137–145
Creaven PJ, Pendyala L, Perez R, Meropol NJ, Loewen GM, Levine E, Berghorn E, Raghavan D (1995) Phase I study of N1N11diethylnorsperimine a polyamine analog in advanced cancer. Proc Am Assoc Clin Oncol 14:472
Faessel HM, Slocum HK, Jackson RC, Boritzki TJ, Rustum YM, Nair MG, Greco WR (1998) Super in vitro synergy between inhibitors of dihydrofolate reductase and inhibitors of other folate-requiring enzymes: the critical role of polyglutamylation. Cancer Res 58:3036–3050
Faessel HM, Slocum HK, Rustum YM, Greco WR (1999) Folic acid-enhanced synergy for the combination of trimetrexate plus the glycinamide ribonucleotide formyltransferase inhibitor 4-[2-(2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4,6][1,4]thiazin-6-yl)-(S)-ethyl]-2,5-thienoylamino-L-glutamic acid (AG2034): comparison across sensitive and resistant human tumor cell lines. Biochem Pharmacol 57:567–577
Fakih MG, Rajput A, Yang GY, Pendyala L, Toth K, Smith JL, Lawrence DD, Rustum YM (2006) A Phase I study of weekly intravenous oxaliplatin in combination with oral daily capecitabine and radiation therapy in the neoadjuvant treatment of rectal adenocarcinoma. Int J Radiat Oncol Biol Phys 65:1462–1470
Feuerstein BG, Williams LD, Basu HS, Marton LJ (1991) Implications and concepts of polyamine-nucleic acid interactions. [Review] [60 refs]. J Cell Biochem 46:37–47
Gessner PK (1974) A straight forward method for the study of drug interactions. In: Morselli PI, Garattini S, Cohen SN (eds) Drug interactions. Raven Press, New York, pp 349–362
Hahm HA, Ettinger DS, Bowling K, Hoker B, Chen TL, Zabelina Y, Casero RAJ (2002) Phase I study of N1,N11-diethylnorspermine in patients with non-small cell lung cancer. Clin Cancer Res 8:684–690
Hawthorne TR, Austin JKJ (1996) Synergism of the polyamine analogue, N1,N11-bisethylnorspermine with cis-diaminedichloroplatinum (II) against murine neoplastic cell lines in vitro and in vivo. Cancer Lett 99:99–107
Hector S, Hawthorn L, Greco W, Pendyala L (2002) Gene expression profiles after oxaliplatin treatment in A2780 ovarian carcinoma cells. Proc Am Assoc Cancer Res 43:62
Hector S, Porter CW, Kramer DL, Clark K, Prey J, Kiesel N, Diegelman P, Chen Y, Pendyala L (2004) Polyamine catabolism in platinum drug action: interactions between oxaliplatin and the polyamine analogue N 1 , N 11-diethylnorspermine at the level of spermidine/spermine N 1-acetylransferase. Mol Cancer Ther 3:813–822
Hector S, Porter CW, Kramer DL, Prey J, Pendyala L (2005) Spermidine/spermine N1-acetyltransferase as a determinant of oxaliplatin drug action and resistance. Proc Am Assoc Cancer Res 46:323
Hector S, Bolanowska-Higdon W, Zdanowicz J, Hitt S, Pendyala L (2001) In vitro studies on the mechanisms of oxaliplatin resistance. Cancer Chemother Pharmacol 48:398–406
Ichimura S, Menoi M, Mita K, Fukuchi K, Hamana K (2004) Accumulation of spermidine/spermine N1-acetyltransferase and alternatively spliced mRNA as a delayed response of HeLa S3 cells following X-ray irradiation. Int J Radiat Biol 80:369–375
Johnson MR, Wang KS, Smith JB, Heslin MJ, Diasio RB (2000) Quantitation of dihydropyrimidine dehydrogenase expression by real-time reverse transcription polymerase chain reaction. Anal Biochem 278:175–184
Maxwell P, Longley DB, Latif T, Boyer J, Allen W, Lynch M, McDermott U, Harkin D, Allegra CJ, Johnston PG (2003) Identification of 5-fluorouracil-inducible target genes using cDNA microarray profiling. Can Res 63:4602–4606
Minchin RF, Knight S, Arulpragasam A, Fogel-Petrovic M (2006) Concentration-dependent effects of N1, N11-diethylnorspermine on melanoma cell proliferation. Int J Cancer 118:509–512
Paliwal J, Janumpalli G, Basu HS (1998) The mechanism of polyamine analog-induced enhancement of cisplatin cytotoxicity in the U-251 MG human malignant glioma cell line. Cancer Chemother Pharmacol 41:398–402
Parry L, Balana FR, Pegg AE (1995) Post-transcriptional regulation of the content of spermidine/spermine N1-acetyltransferase by N1N12-bis(ethyl)spermine. Biochem J 305(Pt 2):451–458
Pizzorno G, Diasio RB, Cheng Y-C (2006) Pyrimidine and purine antimetabolites. In: Kufe DW, Bast RC Jr, Hait WN, Hong WK, Pollock RE, Weichselbaum RC, Holland JF, Frei E III (eds) Cancer medicine. 7. BC Decker, London, pp 661–674
Pledgie A, Huang Y, Hacker A, Zhang Z, Woster PM, Davidson NE, Casero RAJ (2002) 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 280:39843–39851
Porter C, Herrera-Omelas L, Pera P, Petrelli NF, Mittleman A (1987) Polyamine biosynthetic activity in normal and neoplastic human colorectal tissues. Cancer 60:1275–1281
Rixie O, Ortuzar W, Alvarez M, Parker R, Reed E, Paull K, Fojo T (1996) Oxaliplatin, tetraplatin, cisplatin and carboplatin: Spectrum of activity in drug-resistant cell lines and in the cell lines of the National Cancer Institute’s anticancer drug screen panel. Biochem Pharmacol 52:1855–1865
Streiff RR, Bender JF (2001) Phase 1 study of N1-N11-diethylnorspermine (DENSPM) administered TID for 6 days in patients with advanced malignancies. Invest New Drugs 19:29–39
Thomas T, Thomas TJ (2001) Polyamines in cell growth and cell death: molecular mechanisms and therapeutic applications. Cell Mol Life Sci 58:244–258
Tummala R, Porter CW, Diegelman P, Vujcic S, Clark K, Prey J, Kisiel N, Kramer DL, Pendyala L (2007) Platinum drug effects on polyamine enzymes in A2780 human ovarian carcinoma cells. Proc Am Assoc Cancer Res 48:365
Varma R, Hector S, Greco WR, Clark K, Hawthorn L, Porter C, Pendyala L (2007) Platinum drug effects on the expression of genes in the polyamine pathway: time-course and concentration-effect analysis based on Affymetrix gene expression profiling of A2780 ovarian carcinoma cells. Cancer Chemother Pharmacol 59:711–723
Vujcic S, Diegelman P, Bacchi CJ, Kramer DL, Porter CW (2002) Identification and characterization of a novel flavin-containing spermine oxidase of mammalian cell origin. Biochem J 367:665–675
Vujcic S, Halmekyto M, Diegelman P, Gan G, Kramer DL, Janne J, Porter CW (2000) Effects of conditional overexpression of spermidine/spermine N1-acetyltransferase on polyamine pool dynamics, cell growth, and sensitivity to polyamine analogs. J Biol Chem 275:38319–38328
Wiseman LR, Adkins JC, Plosker GL, Goa K (1999) Oxaliplatin. A review of its use in the management of metastatic colorectal cancer. Drugs Aging 14:459–475
Wolff AC, Armstrong DK, Fetting JH, Carducci MK, Riley CD, Bender JF, Casero RAJ, Davidson NE (2001) A Phase II study of the polyamine analog N1,N11-diethylnorspermine (DENSpm) daily for five days every 21 days in patients with previously treated metastatic breast cancer. Clin Cancer Res 9:5922–5928
Zhang W, Ramdas L, Shen W, Song SW, Hu L, Hamilton SR (2003) Apoptotic response to 5-fluorouracil treatment is mediated by reduced polyamines, non-autocrine Fas ligand and induced tumor necrosis factor receptor 2. Cancer Biol Ther 2:572–578
Acknowledgments
This work was supported in part by NCICA109619. Gene expression analysis in tumor biopsies was carried out through other sources. We acknowledge the support of NCI Comprehensive Cancer Center grant CA10656 for the Core Facility usage.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hector, S., Tummala, R., Kisiel, N.D. et al. Polyamine catabolism in colorectal cancer cells following treatment with oxaliplatin, 5-fluorouracil and N 1 , N 11 diethylnorspermine. Cancer Chemother Pharmacol 62, 517–527 (2008). https://doi.org/10.1007/s00280-007-0633-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00280-007-0633-2