Received for publication July 5, 2006.
Revised October 16, 2006.
Accepted for publication October 17, 2006.

Molecular basis for the antiproliferative effect of agmatine in tumor cells of colonic, hepatic and neuronal origin

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 PC12. 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, 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 L-NAME, 1 µM 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 neither due to an interaction with the NO synthases nor with the hypusination of EIF5A nor to 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 investigated agmatine-induced inhibition of cell proliferation was paralleled by an increase in intracellular caspase-3-activity indicating a promotion of apoptosis.

Key words: Nitric oxide synthases, Mechanisms of cell killing/apoptosis, Protein synthesis inhibitors, Tumor suppressors