Abstract
The aim of the present study was to challenge potential mechanisms of action underlying the inhibition of tumor cell proliferation by agmatine. Agmatine inhibited proliferation of the human hepatoma cells HepG2, the human adenocarcinoma cells HT29, the rat hepatoma cells McRH7777, and the rat pheochromocytoma cells PC-12. Inhibition of proliferation of HepG2 cells was associated with an abolition of expression of ornithine decarboxylase (ODC) protein and a doubling of mRNA content encoding ODC. In HepG2 cells, silencing of ODC-antizyme-1, but not of antizyme inhibitor, by RNA interference resulted in an increase of agmatine's antiproliferative effect. Thus, the distinct decrease in intracellular polyamine content by agmatine was due to a reduced translation of the synthesizing protein ODC but was not essentially mediated by induction of ODC-antizyme or blockade of antizyme inhibitor. In interaction experiments 1 mM l-arginine, 1 mM d-arginine, 1 mM citrulline, 100 μM Nω-nitro-l-arginine methyl ester, 1 and 10 μM sodium nitroprusside, and 1 μM N1-guanyl-1,7-diaminoheptane failed to alter agmatine's antiproliferative effect. Hence, the antiproliferative effect of agmatine in HT29 and HepG2 cells is due to an interaction with neither the NO synthases, the hypusination of eIF5A, nor an agmatine-induced reduction in availability of intracellular l-arginine. l-Arginine and citrulline, but not d-arginine, inhibited tumor cell proliferation by themselves. Their inhibitory effect was abolished after silencing of arginine decarboxylase (ADC) expression by RNA interference indicating the conversion to agmatine by ADC. Finally, in the four cell lines under study, agmatine-induced inhibition of cell proliferation was paralleled by an increase in intracellular caspase-3 activity, indicating a promotion of apoptosis.
Footnotes
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This study was supported by grants of the Deutsche Krebshilfe and the Doktor-Robert-Pfleger-Stiftung.
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A preliminary account of the present results was given at the 47th Spring Meeting of the Deutsche Gesellschaft für experimentelle und klinische Pharmakologie und Toxikologie; April 4-6, 2006; Mainz, Germany and in Wolf C, Brüss M, Göthert M, and Molderings GJ (2006) Molecular basis for the antiproliferative action of agmatine (decarboxylated L-arginine). NaunynSchmiedeberg′s Arch Pharmacol372 (Suppl 1):R27.
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ABBREVIATIONS: ODC, ornithine decarboxylase; ADC, arginine decarboxylase; AZI, antizyme inhibitor; eIF5A, eukaryotic translation initiation factor 5A; GC7, N1-guanyl-1,7-diaminoheptane; l-NAME, Nω-nitro-l-arginine methyl ester; ODC-Az, ornithine decarboxylase antizyme-1; PCR, polymerase chain reaction; siRNA, small interfering RNA.
- Received July 4, 2006.
- Accepted October 17, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
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