Gardini Giulia, Cravanzola Carlo, Autelli Riccardo, Testore Giovanni, Cesa Roberta, Morando Laura, Solinas Sandro Paolo, Muzio Giuliana, Grillo Maria Angelica, Colombatto Sebastiano
Dipartimento di Medicina e Oncologia Sperimentale, Sezione di Biochimica, Università di Torino, Via Michelangelo 27, 10126 Turin, Italy.
J Hepatol. 2003 Nov;39(5):793-9. doi: 10.1016/s0168-8278(03)00386-6.
BACKGROUND/AIMS: Previous experiments have shown that agmatine, the product of arginine decarboxylase, is transported in competition with putrescine into quiescent rat hepatocytes, where it promotes several effects, including marked decrease of intracellular polyamines and induction of apoptosis. The primary aim of the present study was to assess the action of agmatine on transformed and proliferating hepatic rat cells.
To assess the effect of agmatine on hepatoma cells, analysis by flow cytometry, Western blotting, reverse transcription-polymerase chain reaction, scanning and transmission electron microscopy, immunofluorescence detection of beta-actin and alpha-tubulin were performed.
The results showed that agmatine has antiproliferative effects on the cell lines studied (HTC, JM2, HepG2). Further experiments were performed on HTC cells. The effect was proportional to agmatine concentration (in a range between 50 and 500 microM). It was not correlated with induction of necrosis or apoptosis and was accompanied by accumulation in G(2)/M cell cycle phase and by dramatic modification of cell morphology. Spermidine reversed these effects, suggesting that the marked decrease of the polyamine pool is the main target of agmatine .
The results obtained show a relationship between the decrease of intracellular polyamine content, the rate of cell growth and the cytoskeleton organization.
背景/目的：先前的实验表明，精氨酸脱羧酶的产物胍丁胺与腐胺竞争性转运进入静止的大鼠肝细胞，在肝细胞中它可产生多种效应，包括显著降低细胞内多胺水平并诱导细胞凋亡。本研究的主要目的是评估胍丁胺对转化和增殖的大鼠肝细胞的作用。
为评估胍丁胺对肝癌细胞的作用，进行了流式细胞术分析、蛋白质免疫印迹法、逆转录-聚合酶链反应、扫描和透射电子显微镜检查以及β-肌动蛋白和α-微管蛋白的免疫荧光检测。
结果显示胍丁胺对所研究的细胞系（HTC、JM2、HepG2）具有抗增殖作用。对HTC细胞进行了进一步实验。该作用与胍丁胺浓度成正比（在50至500微摩尔范围内）。它与坏死或凋亡的诱导无关，且伴随着细胞在G(2)/M细胞周期阶段的积累以及细胞形态的显著改变。亚精胺可逆转这些效应，表明多胺池的显著降低是胍丁胺的主要作用靶点。
所获得的结果表明细胞内多胺含量的降低、细胞生长速率与细胞骨架组织之间存在关联。
