Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1992 Aug 15;286(Pt 1):305–312. doi: 10.1042/bj2860305

Eicosapentaenoic acid inhibits cell growth and triacylglycerol secretion in McA-RH7777 rat hepatoma cultures.

J C Fox 1, R V Hay 1
PMCID: PMC1133055  PMID: 1520281

Abstract

The plasma triacylglycerol-decreasing effect of fish-oil fatty acids was studied in vitro by using the rapidly growing cultured rat hepatoma cell line McA-RH7777. Cells were exposed to albumin-complexed eicosapentaenoic acid (C20:5n-3; EPA), to oleic acid (C18:1n-9; OA), or to albumin alone. Cell growth was similar in albumin- and OA-supplemented cultures, but EPA treatment inhibited growth. As estimated by [14C]glycerol incorporation, OA stimulated both net triacylglycerol synthesis and secretion over control levels in a dose-dependent manner. EPA stimulated triacylglycerol synthesis in similar fashion to OA, but paradoxically decreased net triacylglycerol secretion and led to exaggerated intracellular accumulation of radiolabelled triacylglycerol. The EPA and OA effects were additive at low concentrations of total fatty acid, but at higher fatty acid concentrations OA appeared to negate some effects of EPA. Chemical analysis of albumin- and OA-treated cultures revealed OA-dominant profiles for both cellular and medium triacylglycerol-associated fatty acids. In contrast, EPA was the principal fatty acid in cellular triacylglycerol of EPA-supplemented cultures, whereas medium triacylglycerol from these cultures contained very little EPA. We conclude that McA-RH7777 hepatoma cells readily synthesize EPA-containing triacylglycerol molecules, but they have variable capacity for secreting them. We consider potential mechanisms to account for the effects of EPA in this system.

Full text

PDF
308

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Applegate K. R., Glomset J. A. Effect of acyl chain unsaturation on the conformation of model diacylglycerols: a computer modeling study. J Lipid Res. 1991 Oct;32(10):1635–1644. [PubMed] [Google Scholar]
  2. Applegate K. R., Glomset J. A. Effect of acyl chain unsaturation on the packing of model diacylglycerols in simulated monolayers. J Lipid Res. 1991 Oct;32(10):1645–1655. [PubMed] [Google Scholar]
  3. Axelrod L. Omega-3 fatty acids in diabetes mellitus. Gift from the sea? Diabetes. 1989 May;38(5):539–543. [PubMed] [Google Scholar]
  4. Benner K. G., Sasaki A., Gowen D. R., Weaver A., Connor W. E. The differential effect of eicosapentaenoic acid and oleic acid on lipid synthesis and VLDL secretion in rabbit hepatocytes. Lipids. 1990 Sep;25(9):534–540. doi: 10.1007/BF02537160. [DOI] [PubMed] [Google Scholar]
  5. Black D. D., Hay R. V., Rohwer-Nutter P. L., Ellinas H., Stephens J. K., Sherman H., Teng B. B., Whitington P. F., Davidson N. O. Intestinal and hepatic apolipoprotein B gene expression in abetalipoproteinemia. Gastroenterology. 1991 Aug;101(2):520–528. doi: 10.1016/0016-5085(91)90033-h. [DOI] [PubMed] [Google Scholar]
  6. Chen R. F. Removal of fatty acids from serum albumin by charcoal treatment. J Biol Chem. 1967 Jan 25;242(2):173–181. [PubMed] [Google Scholar]
  7. Daggy B., Arost C., Bensadoun A. Dietary fish oil decreases VLDL production rates. Biochim Biophys Acta. 1987 Aug 15;920(3):293–300. doi: 10.1016/0005-2760(87)90107-x. [DOI] [PubMed] [Google Scholar]
  8. Dallongeville J., Boulet L., Davignon J., Lussier-Cacan S. Fish oil supplementation reduces beta-very low density lipoprotein in type III dysbetalipoproteinemia. Arterioscler Thromb. 1991 Jul-Aug;11(4):864–871. doi: 10.1161/01.atv.11.4.864. [DOI] [PubMed] [Google Scholar]
  9. Davidson B. C., Girao L. A., Giangregorio A., Murphy J. Polyunsaturated fatty acids modulate fibroblast growth in culture. Anticancer Res. 1991 Jan-Feb;11(1):267–272. [PubMed] [Google Scholar]
  10. Donadio J. V., Jr Omega-3 polyunsaturated fatty acids: a potential new treatment of immune renal disease. Mayo Clin Proc. 1991 Oct;66(10):1018–1028. doi: 10.1016/s0025-6196(12)61725-x. [DOI] [PubMed] [Google Scholar]
  11. Dousset N., Negre-Salvayre A., Lopez M., Salvayre R., Douste-Blazy L. Ultraviolet-treated lipoproteins as a model system for the study of the biological effects of lipid peroxides on cultured cell. I. Chemical modifications of ultraviolet-treated low-density lipoproteins. Biochim Biophys Acta. 1990 Aug 6;1045(3):219–223. doi: 10.1016/0005-2760(90)90123-f. [DOI] [PubMed] [Google Scholar]
  12. Ehringer W., Belcher D., Wassall S. R., Stillwell W. A comparison of the effects of linolenic (18:3 omega 3) and docosahexaenoic (22:6 omega 3) acids on phospholipid bilayers. Chem Phys Lipids. 1990 May;54(2):79–88. doi: 10.1016/0009-3084(90)90063-w. [DOI] [PubMed] [Google Scholar]
  13. Elliott W. J., Morrison A. R., Sprecher H. W., Needleman P. The metabolic transformations of columbinic acid and the effect of topical application of the major metabolites on rat skin. J Biol Chem. 1985 Jan 25;260(2):987–992. [PubMed] [Google Scholar]
  14. Ellsworth J. L., Erickson S. K., Cooper A. D. Very low and low density lipoprotein synthesis and secretion by the human hepatoma cell line Hep-G2: effects of free fatty acid. J Lipid Res. 1986 Aug;27(8):858–874. [PubMed] [Google Scholar]
  15. Fernandes G., Venkatraman J. T. Modulation of breast cancer growth in nude mice by omega 3 lipids. World Rev Nutr Diet. 1991;66:488–503. doi: 10.1159/000419316. [DOI] [PubMed] [Google Scholar]
  16. Fisher A. B., Dodia C., Tan Z. T., Ayene I., Eckenhoff R. G. Oxygen-dependent lipid peroxidation during lung ischemia. J Clin Invest. 1991 Aug;88(2):674–679. doi: 10.1172/JCI115352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Garg M. L., Thomson A. B., Clandinin M. T. Hypotriglyceridemic effect of dietary n - 3 fatty acids in rats fed low versus high levels of linoleic acid. Biochim Biophys Acta. 1989 Nov 6;1006(1):127–130. doi: 10.1016/0005-2760(89)90333-0. [DOI] [PubMed] [Google Scholar]
  18. Garg M. L., Wierzbicki A. A., Thomson A. B., Clandinin M. T. Dietary cholesterol and/or n-3 fatty acid modulate delta 9-desaturase activity in rat liver microsomes. Biochim Biophys Acta. 1988 Oct 14;962(3):330–336. doi: 10.1016/0005-2760(88)90262-7. [DOI] [PubMed] [Google Scholar]
  19. Gibbons G. F. Assembly and secretion of hepatic very-low-density lipoprotein. Biochem J. 1990 May 15;268(1):1–13. doi: 10.1042/bj2680001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Goren A., Stankiewicz H., Goldstein R., Drukker A. Fish oil treatment of hyperlipidemia in children and adolescents receiving renal replacement therapy. Pediatrics. 1991 Aug;88(2):265–268. [PubMed] [Google Scholar]
  21. Graham A., Zammit V. A., Brindley D. N. Fatty acid specificity for the synthesis of triacylglycerol and phosphatidylcholine and for the secretion of very-low-density lipoproteins and lysophosphatidylcholine by cultures of rat hepatocytes. Biochem J. 1988 Feb 1;249(3):727–733. doi: 10.1042/bj2490727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Grønn M., Christensen E., Hagve T. A., Christophersen B. O. Peroxisomal retroconversion of docosahexaenoic acid (22:6(n-3)) to eicosapentaenoic acid (20:5(n-3)) studied in isolated rat liver cells. Biochim Biophys Acta. 1991 Jan 4;1081(1):85–91. doi: 10.1016/0005-2760(91)90254-f. [DOI] [PubMed] [Google Scholar]
  23. Harris W. S., Connor W. E., Inkeles S. B., Illingworth D. R. Dietary omega-3 fatty acids prevent carbohydrate-induced hypertriglyceridemia. Metabolism. 1984 Nov;33(11):1016–1019. doi: 10.1016/0026-0495(84)90230-0. [DOI] [PubMed] [Google Scholar]
  24. Harris W. S., Connor W. E., McMurry M. P. The comparative reductions of the plasma lipids and lipoproteins by dietary polyunsaturated fats: salmon oil versus vegetable oils. Metabolism. 1983 Feb;32(2):179–184. doi: 10.1016/0026-0495(83)90226-3. [DOI] [PubMed] [Google Scholar]
  25. Harris W. S. Fish oils and plasma lipid and lipoprotein metabolism in humans: a critical review. J Lipid Res. 1989 Jun;30(6):785–807. [PubMed] [Google Scholar]
  26. Hay R. V., Fleming R. M., Ryan J. W., Williams K. A., Stark V. J., Lathrop K. A., Harper P. V. Nuclear imaging analysis of human low-density lipoprotein biodistribution in rabbits and monkeys. J Nucl Med. 1991 Jun;32(6):1239–1245. [PubMed] [Google Scholar]
  27. Hay R., Fleming R., O'Connell W., Kirschner J., Oppliger W. Apolipoproteins of the orotic acid fatty liver: implications for the biogenesis of plasma lipoproteins. J Lipid Res. 1988 Aug;29(8):981–995. [PubMed] [Google Scholar]
  28. Herold P. M., Kinsella J. E. Fish oil consumption and decreased risk of cardiovascular disease: a comparison of findings from animal and human feeding trials. Am J Clin Nutr. 1986 Apr;43(4):566–598. doi: 10.1093/ajcn/43.4.566. [DOI] [PubMed] [Google Scholar]
  29. Homan R., Grossman J. E., Pownall H. J. Differential effects of eicosapentaenoic acid and oleic acid on lipid synthesis and secretion by HepG2 cells. J Lipid Res. 1991 Feb;32(2):231–241. [PubMed] [Google Scholar]
  30. Hubbard W. C., Hough A. J., Jr, Brash A. R., Watson J. T., Oates J. A. Metabolism of linoleic and arachidonic acids in VX2 carcinoma tissue: identification of monohydroxy octadecadienoic acids and monohydroxy eicosatetraenoic acids. Prostaglandins. 1980 Sep;20(3):431–447. doi: 10.1016/0090-6980(80)90031-3. [DOI] [PubMed] [Google Scholar]
  31. Kanayasu T., Morita I., Nakao-Hayashi J., Asuwa N., Fujisawa C., Ishii T., Ito H., Murota S. Eicosapentaenoic acid inhibits tube formation of vascular endothelial cells in vitro. Lipids. 1991 Apr;26(4):271–276. doi: 10.1007/BF02537136. [DOI] [PubMed] [Google Scholar]
  32. Kinsella J. E., Lokesh B., Stone R. A. Dietary n-3 polyunsaturated fatty acids and amelioration of cardiovascular disease: possible mechanisms. Am J Clin Nutr. 1990 Jul;52(1):1–28. doi: 10.1093/ajcn/52.1.1. [DOI] [PubMed] [Google Scholar]
  33. Kojima T., Terano T., Tanabe E., Okamoto S., Tamura Y., Yoshida S. Long-term administration of highly purified eicosapentaenoic acid provides improvement of psoriasis. Dermatologica. 1991;182(4):225–230. doi: 10.1159/000247800. [DOI] [PubMed] [Google Scholar]
  34. Kromhout D., Bosschieter E. B., de Lezenne Coulander C. The inverse relation between fish consumption and 20-year mortality from coronary heart disease. N Engl J Med. 1985 May 9;312(19):1205–1209. doi: 10.1056/NEJM198505093121901. [DOI] [PubMed] [Google Scholar]
  35. Kromhout D. N-3 fatty acids and coronary heart disease: epidemiology from Eskimos to Western populations. J Intern Med Suppl. 1989;731:47–51. doi: 10.1111/j.1365-2796.1989.tb01435.x. [DOI] [PubMed] [Google Scholar]
  36. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  37. Landymore R. W., Manku M. S., Tan M., MacAulay M. A., Sheridan B. Effects of low-dose marine oils on intimal hyperplasia in autologous vein grafts. J Thorac Cardiovasc Surg. 1989 Nov;98(5 Pt 1):788–791. [PubMed] [Google Scholar]
  38. Lanford R. E., Carey K. D., Estlack L. E., Smith G. C., Hay R. V. Analysis of plasma protein and lipoprotein synthesis in long-term primary cultures of baboon hepatocytes maintained in serum-free medium. In Vitro Cell Dev Biol. 1989 Feb;25(2):174–182. doi: 10.1007/BF02626175. [DOI] [PubMed] [Google Scholar]
  39. Lang C. A., Davis R. A. Fish oil fatty acids impair VLDL assembly and/or secretion by cultured rat hepatocytes. J Lipid Res. 1990 Nov;31(11):2079–2086. [PubMed] [Google Scholar]
  40. Lin D. S., Conner W. E. Are the n-3 fatty acids from dietary fish oil deposited in the triglyceride stores of adipose tissue? Am J Clin Nutr. 1990 Apr;51(4):535–539. doi: 10.1093/ajcn/51.4.535. [DOI] [PubMed] [Google Scholar]
  41. Martin L. J., Reaidi G. B., Gavino G. R., Gavino V. C. Effect of 4,7,10,13,16,19-docosahexaenoic acid on triglyceride accumulation and secretion in rat hepatocytes in culture. Lipids. 1991 May;26(5):374–380. doi: 10.1007/BF02537202. [DOI] [PubMed] [Google Scholar]
  42. Mitaka T., Sattler C. A., Sattler G. L., Sargent L. M., Pitot H. C. Multiple cell cycles occur in rat hepatocytes cultured in the presence of nicotinamide and epidermal growth factor. Hepatology. 1991 Jan;13(1):21–30. [PubMed] [Google Scholar]
  43. Mitaka T., Sattler G. L., Pitot H. C. Amino acid-rich medium (Leibovitz L-15) enhances and prolongs proliferation of primary cultured rat hepatocytes in the absence of serum. J Cell Physiol. 1991 Jun;147(3):495–504. doi: 10.1002/jcp.1041470316. [DOI] [PubMed] [Google Scholar]
  44. Murthy S., Albright E., Mathur S. N., Field F. J. Effect of eicosapentaenoic acid on triacylglycerol transport in CaCo-2 cells. Biochim Biophys Acta. 1990 Jul 16;1045(2):147–155. doi: 10.1016/0005-2760(90)90144-m. [DOI] [PubMed] [Google Scholar]
  45. Narayan K. A., Morris H. P. In vitro synthesis of rat serum lipoproteins and proteins by Morris hepatoma 7777. FEBS Lett. 1972 Nov 1;27(2):311–315. [PubMed] [Google Scholar]
  46. Nestel P. J., Connor W. E., Reardon M. F., Connor S., Wong S., Boston R. Suppression by diets rich in fish oil of very low density lipoprotein production in man. J Clin Invest. 1984 Jul;74(1):82–89. doi: 10.1172/JCI111422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Nestel P. J. Effects of N-3 fatty acids on lipid metabolism. Annu Rev Nutr. 1990;10:149–167. doi: 10.1146/annurev.nu.10.070190.001053. [DOI] [PubMed] [Google Scholar]
  48. Nossen J. O., Rustan A. C., Gloppestad S. H., Målbakken S., Drevon C. A. Eicosapentaenoic acid inhibits synthesis and secretion of triacylglycerols by cultured rat hepatocytes. Biochim Biophys Acta. 1986 Oct 24;879(1):56–65. doi: 10.1016/0005-2760(86)90266-3. [DOI] [PubMed] [Google Scholar]
  49. O'Connor T. P., Roebuck B. D., Peterson F. J., Lokesh B., Kinsella J. E., Campbell T. C. Effect of dietary omega-3 and omega-6 fatty acids on development of azaserine-induced preneoplastic lesions in rat pancreas. J Natl Cancer Inst. 1989 Jun 7;81(11):858–863. doi: 10.1093/jnci/81.11.858. [DOI] [PubMed] [Google Scholar]
  50. Otto D. A., Tsai C. E., Baltzell J. K., Wooten J. T. Apparent inhibition of hepatic triacylglycerol secretion, independent of synthesis, in high-fat fish oil-fed rats: role for insulin. Biochim Biophys Acta. 1991 Feb 26;1082(1):37–48. doi: 10.1016/0005-2760(91)90297-u. [DOI] [PubMed] [Google Scholar]
  51. Parks J. S., Johnson F. L., Wilson M. D., Rudel L. L. Effect of fish oil diet on hepatic lipid metabolism in nonhuman primates: lowering of secretion of hepatic triglyceride but not apoB. J Lipid Res. 1990 Mar;31(3):455–466. [PubMed] [Google Scholar]
  52. Parks J. S., Rudel L. L. Effect of fish oil on atherosclerosis and lipoprotein metabolism. Atherosclerosis. 1990 Oct;84(2-3):83–94. doi: 10.1016/0021-9150(90)90077-v. [DOI] [PubMed] [Google Scholar]
  53. Phillipson B. E., Rothrock D. W., Connor W. E., Harris W. S., Illingworth D. R. Reduction of plasma lipids, lipoproteins, and apoproteins by dietary fish oils in patients with hypertriglyceridemia. N Engl J Med. 1985 May 9;312(19):1210–1216. doi: 10.1056/NEJM198505093121902. [DOI] [PubMed] [Google Scholar]
  54. Rustan A. C., Nossen J. O., Christiansen E. N., Drevon C. A. Eicosapentaenoic acid reduces hepatic synthesis and secretion of triacylglycerol by decreasing the activity of acyl-coenzyme A:1,2-diacylglycerol acyltransferase. J Lipid Res. 1988 Nov;29(11):1417–1426. [PubMed] [Google Scholar]
  55. Sakaguchi M., Imray C., Davis A., Rowley S., Jones C., Lawson N., Keighley M. R., Baker P. R., Neoptolemos J. P. Effects of dietary N-3 and saturated fats on growth rates of the human colonic cancer cell lines SW-620 and LS 174T in vivo in relation to tissue and plasma lipids. Anticancer Res. 1990 Nov-Dec;10(6):1763–1768. [PubMed] [Google Scholar]
  56. Salomon P., Kornbluth A. A., Janowitz H. D. Treatment of ulcerative colitis with fish oil n--3-omega-fatty acid: an open trial. J Clin Gastroenterol. 1990 Apr;12(2):157–161. doi: 10.1097/00004836-199004000-00009. [DOI] [PubMed] [Google Scholar]
  57. Sanders T. A., Sullivan D. R., Reeve J., Thompson G. R. Triglyceride-lowering effect of marine polyunsaturates in patients with hypertriglyceridemia. Arteriosclerosis. 1985 Sep-Oct;5(5):459–465. doi: 10.1161/01.atv.5.5.459. [DOI] [PubMed] [Google Scholar]
  58. Speizer L. A., Watson M. J., Brunton L. L. Differential effects of omega-3 fish oils on protein kinase activities in vitro. Am J Physiol. 1991 Jul;261(1 Pt 1):E109–E114. doi: 10.1152/ajpendo.1991.261.1.E109. [DOI] [PubMed] [Google Scholar]
  59. Strum-Odin R., Adkins-Finke B., Blake W. L., Phinney S. D., Clarke S. D. Modification of fatty acid composition of membrane phospholipid in hepatocyte monolayer with n-3, n-6 and n-9 fatty acids and its relationship to triacylglycerol production. Biochim Biophys Acta. 1987 Sep 25;921(2):378–391. doi: 10.1016/0005-2760(87)90040-3. [DOI] [PubMed] [Google Scholar]
  60. Tanabe S., Sherman H., Smith L., Yang L. A., Fleming R., Hay R. Biogenesis of plasma lipoproteins in rat hepatoma McA-RH7777: importance of diffusion-mediated events during cell growth. In Vitro Cell Dev Biol. 1989 Dec;25(12):1129–1140. doi: 10.1007/BF02621264. [DOI] [PubMed] [Google Scholar]
  61. Thrift R. N., Forte T. M., Cahoon B. E., Shore V. G. Characterization of lipoproteins produced by the human liver cell line, Hep G2, under defined conditions. J Lipid Res. 1986 Mar;27(3):236–250. [PubMed] [Google Scholar]
  62. Tisdale M. J., Beck S. A. Inhibition of tumour-induced lipolysis in vitro and cachexia and tumour growth in vivo by eicosapentaenoic acid. Biochem Pharmacol. 1991 Jan 1;41(1):103–107. doi: 10.1016/0006-2952(91)90016-x. [DOI] [PubMed] [Google Scholar]
  63. Topping D. L., Trimble R. P., Storer G. B. Failure of insulin to stimulate lipogenesis and triacylglycerol secretion in perfused livers from rats adapted to dietary fish oil. Biochim Biophys Acta. 1987 Mar 11;927(3):423–428. doi: 10.1016/0167-4889(87)90108-x. [DOI] [PubMed] [Google Scholar]
  64. Wan J. M., Istfan N. W., Chu C. C., Blackburn G. L., Bistrian B. R. Comparative effects of omega-3 and omega-6 polyunsaturated fatty acids on protein metabolism in rats bearing the mammary adenocarcinoma. Metabolism. 1991 Jun;40(6):577–584. doi: 10.1016/0026-0495(91)90047-z. [DOI] [PubMed] [Google Scholar]
  65. Wong S. H., Fisher E. A., Marsh J. B. Effects of eicosapentaenoic and docosahexaenoic acids on apoprotein B mRNA and secretion of very low density lipoprotein in HepG2 cells. Arteriosclerosis. 1989 Nov-Dec;9(6):836–841. doi: 10.1161/01.atv.9.6.836. [DOI] [PubMed] [Google Scholar]
  66. Wong S. H., Nestel P. J., Trimble R. P., Storer G. B., Illman R. J., Topping D. L. The adaptive effects of dietary fish and safflower oil on lipid and lipoprotein metabolism in perfused rat liver. Biochim Biophys Acta. 1984 Feb 9;792(2):103–109. doi: 10.1016/0005-2760(84)90209-1. [DOI] [PubMed] [Google Scholar]
  67. Wong S., Nestel P. J. Eicosapentaenoic acid inhibits the secretion of triacylglycerol and of apoprotein B and the binding of LDL in Hep G2 cells. Atherosclerosis. 1987 Apr;64(2-3):139–146. doi: 10.1016/0021-9150(87)90239-5. [DOI] [PubMed] [Google Scholar]
  68. Zhang Z. J., Wilcox H. G., Elam M. B., Castellani L. W., Heimberg M. Metabolism of n-3 polyunsaturated fatty acids by the isolated perfused rat liver. Lipids. 1991 Jul;26(7):504–511. doi: 10.1007/BF02536594. [DOI] [PubMed] [Google Scholar]
  69. de Alaniz M. J., de Gómez Dumm I. N., Brenner R. R. Effect of fasting on delta 5 desaturation activity in rat liver microsomes and HTC cells. Mol Cell Biochem. 1980 Dec 16;33(3):165–170. doi: 10.1007/BF00225290. [DOI] [PubMed] [Google Scholar]
  70. de Néchaud B., Fromont S., Bergès J. Enumeration of alphafetoprotein secreting cells using a reverse hemolytic plaque assay during the growth of Morris hepatoma 7777 in culture. Biochem Biophys Res Commun. 1977 Dec 7;79(3):789–795. doi: 10.1016/0006-291x(77)91181-0. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES