Cytotoxicity of polyamines to Amoeba proteus: role of polyamine oxidase.
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 N1-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 N1-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 (approximately 50%). Mepacrine did not inhibit the enzyme activity at pH 8. Neither monoamine nor diamine oxidase activity (approximately 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.
Schenkel
E
E
Laboratory of Toxicology and Bioanalytical Chemistry, Université Libre de Bruxelles (ULB), Belgium.
Dubois
J G
JG
Helson-Cambier
M
M
Hanocq
M
M
D000563:Amebicides / Q000633:toxicity*; D000656:Amoeba / Q000187:drug effects* / Q000201:enzymology*; D000818:Animals; D002478:Cells, Cultured; D004640:Emetine / Q000633:toxicity; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000502:physiology*; D011073:Polyamines / Q000633:toxicity*; D013095:Spermidine / Q000633:toxicity; D013096:Spermine / Q000633:toxicity; D018675:Toxicity Tests; D000097667:Polyamine Oxidase
D000563:Amebicides; D004640:Emetine; D000587:Oxidoreductases Acting on CH-NH Group Donors; D011073:Polyamines; D013095:Spermidine; D013096:Spermine; D000097667:Polyamine Oxidase