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Cytotoxicity of polyamines to Amoeba proteus: Role of polyamine oxidase

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Abstract

It has been shown that oxidation of polyamines by polyamine oxidases can produce toxic compounds (H2O2, aldehydes, ammonia) and that the polyamine oxidase-polyamine system is implicated, in vitro, in the death of several parasites. Using Amoeba proteus as an in vitro model, we studied the cytotoxicity to these cells of spermine, spermidine, their acetyl derivatives, and their hypothetical precursors. Spermine and N 1-acetylspermine were more toxic than emetine, an amoebicidal reference drug. Spermine presented a short-term toxicity, but a 48-h contact time was necessary for the high toxicity of spermidine. The uptake by Amoeba cells of the different polyamines tested was demonstrated. On the other hand, a high polyamine oxidase activity was identified in Amoeba proteus crude extract. Spermine (theoretical 100%) and N 1-acetylspermine (64%) were the best substrates at pH 9.5, while spermidine, its acetyl derivatives, and putrescine were very poorly oxidized by this enzyme (3–20%). Spermine oxidase activity was inhibited by phenylhydrazine (nil) and isoniazid (≈ 50%). Mepacrine did not inhibit the enzyme activity at pH 8. Neither monoamine nor diamine oxidase activity (≈ 10%) was found. It must be emphasized that spermine, the best enzyme substrate, is the most toxic polyamine. This finding suggests that knowledge of polyamine oxidase specificity can be used to modulate the cytotoxicity of polyamine derivatives. Amoeba proteus was revealed as a simple model for investigation of the connection between cytotoxicity and enzyme activity.

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Abbreviations

DAO:

diamine oxidase

DFMO:

DL-α-difluoromethylornithine

DP:

1-3-diaminopropane

IC50 :

50% inhibition concentration

MAO:

monoamine oxidase

N 1-ACSP;:

N 1-acetylspermine

N1-ACSPD:

N 1-acetylspermidine

N 8-ACSPD:

N 8-acetylspermidine

ODC:

ornithine decarboxylase

PAO(s):

polyamine oxidase(s)

PUT:

putrescine

SP:

spermine

SPD:

spermidine

References

  • Abi Khalil F, Dubois J, Hanocq M, Atassi G. A new algorithm for computing the parameters of linear models in pharmacokinetics. Eur J Drug Metab Pharmacokinet. 1986;11:51–9.

    Google Scholar 

  • Akaïke H. A new look at the statistical model identification. IEEE Trans Autom Contr. 1974;AC19:716–23.

    Google Scholar 

  • Averill-Bates DA, Agostinelli E, Przybytkowski E, Mateescu MA, Mondovi B. Cytotoxicity and kinetic analysis of purified bovine serum amine oxidase in the presence of spermine in chinese hamster ovary cells. Arch Biochem Biophys. 1993;300:75–9.

    Google Scholar 

  • Bacchi CJ, Yarlett N. Effects of antagonists of polyamine metabolism on African trypanosomes. Acta Tropica. 1993;54:225–36.

    Google Scholar 

  • Davis RH. Management of polyamine pools and the regulation of ornithine decarboxylase. J Cell Biochem. 1990;44:199–205.

    Google Scholar 

  • Dubois J, Abi Khalil F, Hanocq M, Atassi G, Arnould R. Ajustement des courbes dose-effet au moyen d'un algorithme original. Applications à des études de cytotoxicité in vitro. J Pharm Belg. 1989;44:181–91.

    Google Scholar 

  • Duez P, Livaditis A, Guissou PI, Sawadogo M, Hanocq M. Use of an Amoeba proteus model for in vitro cytotoxicity testing in phytochemical research. Application to Euphorbia hirta extracts. J Ethnopharm. 1991;34:235–46.

    Google Scholar 

  • Ferrante A, Rzepczyk CM, Saul AJ. Polyamine oxidase-mediated trypanosome killing: the role of hydrogen peroxide and aldehydes. J Immunol. 1984;133:2157–62.

    Google Scholar 

  • Ferrante A, Ljungstrom I, Rzepczyk CM, Morgan DML. Differences in sensitivity of Schistosoma mansoni schistosomula, Dirofilaria immitis microfilariae, and Nematospiroides dubius third-stage larvae to damage by the polyamine oxidase-polyamine system. Infect Immun. 1986;53:606–10.

    Google Scholar 

  • Gramzinski R, Parchment R, Pierce G. Evidence linking programmed cell death in the blastocyst to polyamine oxidase. Differentiation. 1990;43:59–65.

    Google Scholar 

  • Griffin JL. An improved mass culture method for the large free living amoeba. Exp Cell Res. 1960;21:170–8.

    Google Scholar 

  • Haas E. The effect of atabrine and quinine on isolated respiratory enzymes. J Biol Chem. 1944;155:321–31.

    Google Scholar 

  • Heby O, Andersson G. Polyamines and the cell cycle. In: Gaugas JM, ed. Polyamines in biomedical research. Chichester: Wiley; 1980:1–16.

    Google Scholar 

  • Henle KJ, Moss AJ, Nagle WA. Mechanism of spermidine cytotoxicity at 37°C and 43°C in chinese hamster ovary cells. Cancer Res. 1986;46:175–82.

    Google Scholar 

  • Hill AV. The possible effects of the aggregation of the molecules of haemoglobin on its dissociation curves. J Physiol. 1910;40:IV-VIII.

    Google Scholar 

  • Hölttä E. Oxidation of spermidine and spermine in rat liver. Purification and properties of polyamine oxidase. Biochemistry. 1977;16:91–100.

    Google Scholar 

  • Kim B, Sobota A, Bitonti A, McCann P, Byers T. Polyamine metabolism in Acanthamoeba: polyamine content and synthesis of ornithine, putrescine and diaminopropane. Protozool. 1987;34:278–84.

    Google Scholar 

  • Levitz SM, Dibenedetto DJ, Diamond RD. Inhibition and killing of fungi by the polyamine oxidase-polyamine system. Antonie van Leeuwenhoek. 1990;58:107–14.

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–75.

    Google Scholar 

  • Matsumoto T, Nimura Y, Furuta T et al. Some properties of amine oxidase from soybean seedlings. Nagoya J Med Sci. 1984;46:87–94.

    Google Scholar 

  • Morgan DML. Polyamine oxidases. In: Gaugas JM, ed. Polyamines in biomedical research. Chichester: Wiley; 1980: 285–302.

    Google Scholar 

  • Morgan DML. Polyamine oxidases. Biochem Soc Trans. 1985;13:322–5.

    Google Scholar 

  • Morgan DML. Polyamines and cellular regulation: perspectives. Biochem Soc Trans. 1990;18:1080–4.

    Google Scholar 

  • Morris DR, Harada J. Participation of polyamines in the proliferation of bacterial and animal cells. In: Gaugas JM, ed. Polyamines in biomedical research. Chichester: Wiley; 1980:17–34.

    Google Scholar 

  • Nikolov I, Pavlov V, Minkov I, Damjanov D. Purification and some properties of amine oxidase from soybean seedlings. Experientia. 1990;46:765–7.

    Google Scholar 

  • Parchment RE, Lewellyn A, Swartzendruber D, Pierce GB. Serum amine oxidase activity contributes to crisis in mouse embryo cell lines. Proc Natl Acad Sci USA. 1990;87:4340–4.

    Google Scholar 

  • Pegg AE. Recent advances in the biochemistry of polyamines in eukaryotes. Biochem J. 1986;234:249–62.

    Google Scholar 

  • Pegg AE. Polyamine metabolism and its importance in neoplastic growth and as a target for chemotherapy. Cancer Res. 1988;48:759–74.

    Google Scholar 

  • Poulin R, Larochelle J, Nadeau P. Polyamines in Acanthamoeba castellanii: presence of an unusually high, osmotically sensitive pool of 1,3-diaminopropane. Biochem Biophys Res Commun. 1984;122:388–93.

    Google Scholar 

  • Quemener V, Khan NA, Moulinoux JPh. Polyamines et cancer. Cancer J. 1990;3:45–52.

    Google Scholar 

  • Rahiala E-L, Kekomäki M, Jänne J, Raina A, Raina NCR. Inhibition of pyridoxal enzymes by l-canaline. Biochim Biophys Acta. 1971;227:337–43.

    Google Scholar 

  • Rzepczyk CM, Saul AJ, Ferrante A. Polyamine oxidase-mediated intraerythrocytic killing of Plasmodium falciparum: evidence against the role of reactive oxygen metabolites. Infect Immun. 1984;43:238–44.

    Google Scholar 

  • Schenkel E, Berlaimont V, Dubois J, Helso-Cambier M, Hanocq M. Improved HPLC method for the determination of polyamines as their benzoylated derivatives. Application to P388 cancer cells. J Chromatogr. 1995;668:189–97.

    Google Scholar 

  • Shukla OP, Müller S, Walter RD. Polyamine oxidase from Acanthamoeba culbertsoni specific for N 8-acetylspermidine. Mol Biochem Parasitol. 1992;51:91–8.

    Google Scholar 

  • Smith TA. The di- and poly-amine oxidases of higher plants. Biochem Soc Trans. 1985;13:319–21.

    Google Scholar 

  • Tabor CW, Kellog PD. Spermidine deshydrogenase (Serratia marcescens). In: Tabor H, Tabor CW, eds. Methods in enzymology. vol.17B. New York: Academic Press; 1971: 746–53.

    Google Scholar 

  • Tabor CW, Tabor H. Polyamines in microorganisms. Microbiol Rev. 1985;49:81–99.

    Google Scholar 

  • Tabor CW, Tabor H, Bachrach U. Identification of the aminoaldehydes produced by the oxidation of spermine and spermidine with purified plasma amine oxidase. J Biol Chem. 1964;239:2194–203.

    Google Scholar 

  • Yarlett N. Polyamine biosynthesis and inhibition in Trichomonas vaginalis. Parasitol Today. 1988;4:357–60.

    Google Scholar 

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Authors and Affiliations

  1. Laboratory of Toxicology and Bioanalytical Chemistry, Université Libre de Bruxelles (ULB), Institute of Pharmacy, Brussels, Belgium

    E. Schenkel, J. G. Dubois, M. Helson-Cambier & M. Hanocq

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  1. E. Schenkel
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  2. J. G. Dubois
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  3. M. Helson-Cambier
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  4. M. Hanocq
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Schenkel, E., Dubois, J.G., Helson-Cambier, M. et al. Cytotoxicity of polyamines to Amoeba proteus: Role of polyamine oxidase. Cell Biol Toxicol 12, 1–9 (1996). https://doi.org/10.1007/BF00143389

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