Abstract
An occurrence and a magnitude of alcoholic liver diseases depend on the balance between ethanol-induced injury and liver regeneration. Like ethanol, polyamines including putrescine, spermidine, and spermine modulate cell proliferation. Thus, the purpose of this study was to evaluate the relationship between effect of ethanol on hepatocyte (HC) proliferation and polyamine metabolism using the HepaRG cell model. Results showed that ethanol effect in proliferating HepaRG cells was associated with a decrease in intracellular polyamine levels and ornithine decarboxylase (ODC) activity. Ethanol also induced disorders in expression of genes coding for polyamine-metabolizing enzymes. The α-difluoromethyl ornithine, an irreversible inhibitor of ODC, amplified ethanol toxicity on cell viability, protein level, and DNA synthesis through accentuation of polyamine depletion in proliferating HepaRG cells. Conversely, putrescine reversed ethanol effect on cell proliferation parameters. In conclusion, this study suggested that ethanol effect on HC proliferation was closely related to polyamine metabolism and that manipulation of this metabolism by putrescine could protect against the anti-proliferative activity of ethanol.
Similar content being viewed by others
References
Aninat C, Piton A, Glaise D, Le Charpentier T, Langouet S, Morel F, Guguen-Guillouzo C, Guillouzo A (2006) Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells. Drug Metab Dispos 34(1):75–83
Baumgardner JN, Shankar K, Korourian S, Badger TM, Ronis MJ (2007) Undernutrition enhances alcohol-induced hepatocyte proliferation in the liver of rats fed via total enteral nutrition. Am J Physiol Gastrointest Liver Physiol 293(1):G355–G364
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Chen KY, Presepe V, Parken N, Liu AY (1982) Changes of ornithine decarboxylase activity and polyamine content upon differentiation of mouse NB-15 neuroblastoma cells. J Cell Physiol 110(3):285–290
Denizot F, Lang R (1986) Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 89(2):271–277
Diehl AM, Abdo S, Brown N (1990) Supplemental putrescine reverses ethanol-associated inhibition of liver regeneration. Hepatology 12(4 Pt 1):633–637
Diehl AM, Yang SQ, Brown N, Smith J, Raiford D, Gordon R, Casero R (1992) Ethanol-associated alterations in the kinetics of putrescine uptake and metabolism by the regenerating liver. Alcohol Clin Exp Res 16(1):5–10
Do THT, Gaboriau F, Ropert M, Moirand R, Cannie I, Brissot P, Loréal O, Lescoat G (2011) Ethanol effect on cell proliferation in the human hepatoma heparg cell line: relationship with iron metabolism. Alcohol Clin Exp Res 35(3):408–419
Ducros V, Ruffieux D, Belva-Besnet H, de Fraipont F, Berger F, Favier A (2009) Determination of dansylated polyamines in red blood cells by liquid chromatography-tandem mass spectrometry. Anal Biochem 390(1):46–51
Ewton DZ, Erwin BG, Pegg AE, Florini JR (1984) The role of polyamines in somatomedin-stimulated differentiation of L6 myoblasts. J Cell Physiol 120(3):263–270
Gaboriau F, Havouis R, Groussard K, Moulinoux JP, Lescoat G (2005) Measurement of ornithine decarboxylase activity in cell extracts using mass spectrometry detection of dansylated putrescine. Anal Biochem 341(2):385–387
Gerner EW, Mamont PS, Bernhardt A, Siat M (1986) Post-translational modification of the protein-synthesis initiation factor eIF-4D by spermidine in rat hepatoma cells. Biochem J 239(2):379–386
Gripon P, Rumin S, Urban S, Le Seyec J, Glaise D, Cannie I, Guyomard C, Lucas J, Trepo C, Guguen-Guillouzo C (2002) Infection of a human hepatoma cell line by hepatitis B virus. Proc Natl Acad Sci USA 99(24):15655–15660
Gripon P, Cannie I, Urban S (2005) Efficient inhibition of hepatitis B virus infection by acylated peptides derived from the large viral surface protein. J Virol 79(3):1613–1622
Gugliucci A (2004) Polyamines as clinical laboratory tools. Clin Chim Acta 344(1–2):23–35
Jacobs JS, Miller MW (2001) Proliferation and death of cultured fetal neocortical neurons: effects of ethanol on the dynamics of cell growth. J Neurocytol 30(5):391–401
Klein RF, Carlos AS (1995) Inhibition of osteoblastic cell proliferation and ornithine decarboxylase activity by ethanol. Endocrinology 136(8):3406–3411
Koivisto T, Salaspuro M (1998) Acetaldehyde alters proliferation, differentiation and adhesion properties of human colon adenocarcinoma cell line Caco-2. Carcinogenesis 19(11):2031–2036
Koteish A, Yang S, Lin H, Huang J, Diehl AM (2002) Ethanol induces redox-sensitive cell-cycle inhibitors and inhibits liver regeneration after partial hepatectomy. Alcohol Clin Exp Res 26(11):1710–1718
Linsalata M, Giannini R, Notarnicola M, Cavallini A (2006) Peroxisome proliferator-activated receptor gamma and spermidine/spermine N1-acetyltransferase gene expressions are significantly correlated in human colorectal cancer. BMC Cancer 6:191
Liu JL, Du J, Fan LL, Liu XY, Gu L, Ge YB (2008) Effects of quercetin on hyper-proliferation of gastric mucosal cells in rats treated with chronic oral ethanol through the reactive oxygen species-nitric oxide pathway. World J Gastroenterol 14(20):3242–3248
Minuk GY, Gauthier T (1993) The effect of gamma-aminobutyric acid on hepatic regenerative activity following partial hepatectomy in rats. Gastroenterology 104(1):217–221
Norikura T, Kojima-Yuasa A, Opare Kennedy D, Matsui-Yuasa I (2007) Protective effect of gamma-aminobutyric acid (GABA) against cytotoxicity of ethanol in isolated rat hepatocytes involves modulations in cellular polyamine levels. Amino Acids 32(3):419–423
Ramirez-Farias C, Madrigal-Santillan E, Gutierrez-Salinas J, Rodriguez-Sanchez N, Martinez-Cruz M, Valle-Jones I, Gramlich-Martinez I, Hernandez-Ceruelos A, Morales-Gonzalez JA (2008) Protective effect of some vitamins against the toxic action of ethanol on liver regeneration induced by partial hepatectomy in rats. World J Gastroenterol 14(6):899–907
Seiler N (1970) Use of the dansyl reaction in biochemical analysis. Methods Biochem Anal 18:259–337
Seiler N, Raul F (2005) Polyamines and apoptosis. J Cell Mol Med 9(3):623–642
Sessa A, Perin A (1997) Ethanol and polyamine metabolism: physiologic and pathologic implications: a review. Alcohol Clin Exp Res 21(2):318–325
Tanaka T, Kurai K, Kunitoh S, Kondo K, Goto Y, Kawai S, Warashina M, Yamashita T, Toda T, Monna T et al (1993) Gender-related differences in the inhibitory effect on liver regeneration in alcohol-treated rats: study of polyamine metabolism. Alcohol Alcohol Suppl 1A:15–20
Troadec MB, Glaise D, Lamirault G, Le Cunff M, Guerin E, Le Meur N, Detivaud L, Zindy P, Leroyer P, Guisle I, Duval H, Gripon P, Theret N, Boudjema K, Guguen-Guillouzo C, Brissot P, Leger JJ, Loreal O (2006) Hepatocyte iron loading capacity is associated with differentiation and repression of motility in the HepaRG cell line. Genomics 87(1):93–103
Verma DS, Sunkara PS (1982) An essential role for polyamine biosynthesis during human granulopoietic differentiation. Cancer Res 42(8):3046–3049
Vignesh RC, Sitta Djody S, Jayasudha E, Gopalakrishnan V, Ilangovan R, Balaganesh M, Veni S, Sridhar M, Srinivasan N (2006) Effect of ethanol on human osteosarcoma cell proliferation, differentiation and mineralization. Toxicology 220(1):63–70
Wallace HM, Fraser AV, Hughes A (2003) A perspective of polyamine metabolism. Biochem J 376(Pt 1):1–14
Yatin M, Venkataraman GM, Marcinek R, Ain KB (1999) Polyamine synthesis and transport inhibition in a human anaplastic thyroid carcinoma cell line in vitro and as xenograft tumors. Thyroid 9(8):805–814
Acknowledgments
This study was supported by Institut de Recherches Scientifiques sur les Boissons (IREB) and Ligue contre le Cancer (Ille et Vilaine/Loire Atlantique).
Conflict of interest
All authors do not have conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Do, T.H.T., Gaboriau, F., Morel, I. et al. Modulation of ethanol effect on hepatocyte proliferation by polyamines. Amino Acids 44, 869–877 (2013). https://doi.org/10.1007/s00726-012-1413-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-012-1413-9