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Polyamines pp 367-378 | Cite as

Determination of N1,N12-Diacetylspermine in Urine: A Novel Tumor Marker

  • Masao Kawakita
  • Kyoko Hiramatsu
  • Mari Yanagiya
  • Yosuke Doi
  • Mieko Kosaka
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 720)

Abstract

N 1,N 12-diacetylspermine (DiAcSpm) is a minor component of human urine that constitutes less than 0.5% of total polyamine species in human urine. Structurally related polyamines and acetylpolyamines were separated and analyzed by HPLC and gas chromatography, and refinement of these procedures led to the identification of this minor component. Subsequent analyses of urines from cancer patients as well as healthy persons revealed that DiAcSpm is a promising candidate for a novel tumor marker. It is much more sensitive than established tumor markers in detecting colorectal and other cancers, and most importantly, is able to detect 60% of early colorectal cancers confined to mucous membranes. Serum CEA is able to detect only about 10% of colorectal cancers at this stage. Collection of urine is easy and does not give any pain to patients, which adds another merit to urinary DiAcSpm as a tumor marker. DiAcSpm-specific antibodies were then developed for simpler determination of DiAcSpm in urine, and the antibodies were used to construct an ELISA system. More recently, a reagent kit for DiAcSpm determination based on colloidal gold aggregation that can be used with automatic biochemical analyzers was also developed.

Key words

Diacetylspermine Tumor marker Urine Cancer diagnosis Immunoassay Early cancer detection ELISA Automated determination 

References

  1. 1.
    Bachrach U (1992) Polyamines as markers of malignancy. Prog Drug Res 39:9–33PubMedGoogle Scholar
  2. 2.
    Hiramatsu K, Sugimoto M, Kamei S, Hoshino M, Kinoshita K, Iwasaki K, Kawakita M (1995) Determination of the amounts of polyamines excreted in urine: demonstration of N1, N8-diacetylspermidine and N1, N12-diacetylspermine as components commonly occurring in normal human urine. J Biochem 117:107–112PubMedGoogle Scholar
  3. 3.
    Sugimoto M, Hiamatsu K, Kamei S, Kinoshita K, Hoshino M, Iwasaki K, Kawakita M (1995) Significance of urinary N1, N8-diacetylspermidine and N1, N12-diacetylspermine as indicators of neoplastic diseases. J Cancer Res Clin Oncol 121:317–319PubMedCrossRefGoogle Scholar
  4. 4.
    Hiramatsu K, Sugimoto M, Kamei S, Hoshino M, Kinoshita K, Iwasaki K, Kawakita M (1997) Diagnostic and prognostic usefulness of N1, N8-diacetylspermidine and N1, N12-diacetylspermine in urine as novel markers of malignancy. J Cancer Res Clin Oncol 123:539–545PubMedCrossRefGoogle Scholar
  5. 5.
    Hiramatsu K, Miura H, Kamei S, Iwasaki K, Kawakita M (1998) Development of a sensitive and accurate enzyme-linked immunosorbent assay (ELISA) system that can substitute the HPLC analysis for the determination of N1, N12-diacetylspermine in human urine. J Biochem 124:231–236PubMedGoogle Scholar
  6. 6.
    Hiramatsu K, Takahashi K, Yamaguchi T, Matsumoto H, Miyamoto H, Tanaka S, Tanaka C, Tamamori Y, Imajo M, Kawaguchi M, Toi M, Mori T, Kawakita M (2005) N1, N12-Diacetylspermine as a sensitive and specific novel marker for early- and late-stage colorectal and breast cancers. Clin Cancer Res 11:2986–2990PubMedCrossRefGoogle Scholar
  7. 7.
    van den Berg GA, Muskiet F, Kingma AW, van den Silk W, Halie MR (1986) Simultaneous gas-chromatographic determination of free and acetyl-conjugated polyamines in urine. Clin Chem 32:1930–1937PubMedGoogle Scholar
  8. 8.
    Vujcic S, Halmekyto M, Diegelman P, Gan G, Kramer DL, Jänne 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 275:38319–38328PubMedCrossRefGoogle Scholar
  9. 9.
    Hamaoki M, Hiramatsu K, Suzuki S, Nagata A, Kawakita M (2002) Two enzyme-linked immunosorbent assay (ELISA) systems for N1, N8-diacetylspermidine and N1 N12-diacetylspermine using monoclonal antibodies. J Biochem 132:783–788PubMedCrossRefGoogle Scholar
  10. 10.
    Kobayashi M, Samejima K, Hiramatsu K, Kawakita M (2002) Mass spectrometric separation and determination of N1, N12-diacetylspermine in the urine of cancer patients. Biol Pharm Bull 25:372–374PubMedCrossRefGoogle Scholar
  11. 11.
    Hiramatsu K, Miura H, Sugimoto K, Kamei S, Iwasaki K, Kawakita M (1997) Preparation of antibodies highly specific to N1, N8-diacetylspermidine, and development of an enzyme-linked immunosorbent assay (ELISA) system for its sensitive and specific detection. J Biochem 121:1134–1138PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Masao Kawakita
    • 1
  • Kyoko Hiramatsu
    • 1
  • Mari Yanagiya
    • 2
  • Yosuke Doi
    • 2
  • Mieko Kosaka
    • 2
  1. 1.Department of Molecular Medical ResearchTokyo Metropolitan Institute of Medical ScienceSetagaya-kuJapan
  2. 2.Alfresa Pharma CorporationOsakaJapan

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