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. 1993 Sep 15;294(Pt 3):917–921. doi: 10.1042/bj2940917

Hormonal and substrate regulation of 3-thia fatty acid metabolism in Morris 7800 C1 hepatoma cells.

E Hvattum 1, H J Grav 1, J Bremer 1
PMCID: PMC1134549  PMID: 8379945

Abstract

The 3-thia fatty acid tetradecylthioacetic acid (TTA) has recently been shown to inhibit growth rate and increase peroxisomal acyl-CoA oxidase (ACO) (EC 1.3.99.3) activity in the Morris 7800 C1 hepatoma cells. Dexamethasone potentiates and insulin antagonizes these effects of TTA. We demonstrate here the metabolism of the 3-thia acids in these cells and the influence of insulin and dexamethasone on this. (1) The Morris 7800 C1 hepatoma cells exhibited a low omega-hydroxylation activity of the 3-thia acid (and lauric acid). The combination of TTA and dexamethasone induced the omega-hydroxylation and ACO activities in these cells. TTA alone induced ACO activity, but not omega-hydroxylation activity. Insulin counteracted the induction of both enzyme activities. These results indicate that these two enzyme activities are under similar but independent regulation. (2) Hepatoma cells grown with 80 microM TTA in the medium accumulated phospholipids containing the 3-thia fatty acid. After 7 days, TTA accounted for approx. 40% of the total fatty acids in the phospholipids. In addition, TTA affected the incorporation of endogenous fatty acids into phospholipids by decreasing the amounts of palmitic (C16:0) and vaccenic (C18:1(n-7)) acid and increasing the amounts of linoleic (C18:2(n-6)) and alpha-linolenic (C18:3(n-3)) acid in the phospholipids. (3) Dexamethasone increased the incorporation of labelled TTA into both phospholipids and triacylglycerol. Most of the labelled triacylglycerol formed was secreted into the medium. Insulin increased the incorporation of labelled TTA into triacylglycerol, but not into phospholipids. The labelled triacylglycerol formed was retained in the cells.

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  1. Bartlett S. M., Gibbons G. F. Short- and longer-term regulation of very-low-density lipoprotein secretion by insulin, dexamethasone and lipogenic substrates in cultured hepatocytes. A biphasic effect of insulin. Biochem J. 1988 Jan 1;249(1):37–43. doi: 10.1042/bj2490037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell D. R., Elcombe C. R. Induction of acyl-CoA oxidase and cytochrome P450IVA1 RNA in rat primary hepatocyte culture by peroxisome proliferators. Biochem J. 1991 Nov 15;280(Pt 1):249–253. doi: 10.1042/bj2800249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berge R. K., Aarsland A., Kryvi H., Bremer J., Aarsaether N. Alkylthio acetic acids (3-thia fatty acids)--a new group of non-beta-oxidizable peroxisome-inducing fatty acid analogues--II. Dose-response studies on hepatic peroxisomal- and mitochondrial changes and long-chain fatty acid metabolizing enzymes in rats. Biochem Pharmacol. 1989 Nov 15;38(22):3969–3979. doi: 10.1016/0006-2952(89)90676-x. [DOI] [PubMed] [Google Scholar]
  4. Bergseth S., Bremer J. Alkylthioacetic acids (3-thia fatty acids) are metabolized and excreted as shortened dicarboxylic acids in vivo. Biochim Biophys Acta. 1990 May 22;1044(2):237–242. doi: 10.1016/0005-2760(90)90308-k. [DOI] [PubMed] [Google Scholar]
  5. Beynen A. C., Haagsman H. P., Van Golde L. M., Geelen M. J. The effects of insulin and glucagon on the release of triacylglycerols by isolated rat hepatocytes are mere reflections of the hormonal effects on the rate of triacylglycerol synthesis. Biochim Biophys Acta. 1981 Jul 24;665(1):1–7. doi: 10.1016/0005-2760(81)90224-1. [DOI] [PubMed] [Google Scholar]
  6. Chalvardjian A., Rudnicki E. Determination of lipid phosphorus in the nanomolar range. Anal Biochem. 1970 Jul;36(1):225–226. doi: 10.1016/0003-2697(70)90352-0. [DOI] [PubMed] [Google Scholar]
  7. Dich J., Bro B., Grunnet N., Jensen F., Kondrup J. Accumulation of triacylglycerol in cultured rat hepatocytes is increased by ethanol and by insulin and dexamethasone. Biochem J. 1983 Jun 15;212(3):617–623. doi: 10.1042/bj2120617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Duerden J. M., Bartlett S. M., Gibbons G. F. Long-term maintenance of high rates of very-low-density-lipoprotein secretion in hepatocyte cultures. A model for studying the direct effects of insulin and insulin deficiency in vitro. Biochem J. 1989 Nov 1;263(3):937–943. doi: 10.1042/bj2630937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Durrington P. N., Newton R. S., Weinstein D. B., Steinberg D. Effects of insulin and glucose on very low density lipoprotein triglyceride secretion by cultured rat hepatocytes. J Clin Invest. 1982 Jul;70(1):63–73. doi: 10.1172/JCI110604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fox J. C., Hay R. V. Eicosapentaenoic acid inhibits cell growth and triacylglycerol secretion in McA-RH7777 rat hepatoma cultures. Biochem J. 1992 Aug 15;286(Pt 1):305–312. doi: 10.1042/bj2860305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hvattum E., Bergseth S., Pedersen C. N., Bremer J., Aarsland A., Berge R. K. Microsomal oxidation of dodecylthioacetic acid (a 3-thia fatty acid) in rat liver. 1991 Mar 15-Apr 1Biochem Pharmacol. 41(6-7):945–953. doi: 10.1016/0006-2952(91)90200-o. [DOI] [PubMed] [Google Scholar]
  12. Kaluzny M. A., Duncan L. A., Merritt M. V., Epps D. E. Rapid separation of lipid classes in high yield and purity using bonded phase columns. J Lipid Res. 1985 Jan;26(1):135–140. [PubMed] [Google Scholar]
  13. Kusunose E., Ogita K., Ichihara K., Kusunose M. Effect of cytochrome b5 on fatty acid omega- and (omega-1)-hydroxylation catalyzed by partially purified cytochrome P-450 from rabbit kidney cortex microsomes. J Biochem. 1981 Oct;90(4):1069–1076. doi: 10.1093/oxfordjournals.jbchem.a133558. [DOI] [PubMed] [Google Scholar]
  14. Labarca C., Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. doi: 10.1016/0003-2697(80)90165-7. [DOI] [PubMed] [Google Scholar]
  15. Lake B. G., Gray T. J., Pels Rijcken W. R., Beamand J. A., Gangolli S. D. The effect of hypolipidaemic agents on peroxisomal beta-oxidation and mixed-function oxidase activities in primary cultures of rat hepatocytes. Relationship between induction of palmitoyl-CoA oxidation and lauric acid hydroxylation. Xenobiotica. 1984 Mar;14(3):269–276. doi: 10.3109/00498258409151411. [DOI] [PubMed] [Google Scholar]
  16. Lock E. A., Mitchell A. M., Elcombe C. R. Biochemical mechanisms of induction of hepatic peroxisome proliferation. Annu Rev Pharmacol Toxicol. 1989;29:145–163. doi: 10.1146/annurev.pa.29.040189.001045. [DOI] [PubMed] [Google Scholar]
  17. Martin-Sanz P., Vance J. E., Brindley D. N. Stimulation of apolipoprotein secretion in very-low-density and high-density lipoproteins from cultured rat hepatocytes by dexamethasone. Biochem J. 1990 Nov 1;271(3):575–583. doi: 10.1042/bj2710575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nilsson A., Arey H., Pedersen J. I., Christiansen E. N. The effect of high-fat diets on microsomal lauric acid hydroxylation in rat liver. Biochim Biophys Acta. 1986 Nov 14;879(2):209–214. doi: 10.1016/0005-2760(86)90104-9. [DOI] [PubMed] [Google Scholar]
  19. Norrheim L., Sørensen H., Gautvik K., Bremer J., Spydevold O. Synergistic actions of tetradecylthioacetic acid (TTA) and dexamethasone on induction of the peroxisomal beta-oxidation and on growth inhibition of Morris hepatoma cells. Both effects are counteracted by insulin. Biochim Biophys Acta. 1990 Mar 9;1051(3):319–323. doi: 10.1016/0167-4889(90)90141-y. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Ortiz de Montellano P. R., Chan W. K., Tuck S. F., Kaikaus R. M., Bass N. M., Peterson J. A. Mechanism-based probes of the topology and function of fatty acid hydroxylases. FASEB J. 1992 Jan 6;6(2):695–699. doi: 10.1096/fasebj.6.2.1537458. [DOI] [PubMed] [Google Scholar]
  22. Patsch W., Franz S., Schonfeld G. Role of insulin in lipoprotein secretion by cultured rat hepatocytes. J Clin Invest. 1983 May;71(5):1161–1174. doi: 10.1172/JCI110865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Richardson U. I., Snodgrass P. J., Nuzum C. T., Tashjian A. H., Jr Establishment of a clonal strain of hepatoma cells which maintain in culture the five enzymes of the urea cycle. J Cell Physiol. 1974 Feb;83(1):141–149. doi: 10.1002/jcp.1040830117. [DOI] [PubMed] [Google Scholar]
  24. Skrede S., Narce M., Bergseth S., Bremer J. The effects of alkylthioacetic acids (3-thia fatty acids) on fatty acid metabolism in isolated hepatocytes. Biochim Biophys Acta. 1989 Oct 17;1005(3):296–302. doi: 10.1016/0005-2760(89)90052-0. [DOI] [PubMed] [Google Scholar]
  25. Small G. M., Burdett K., Connock M. J. A sensitive spectrophotometric assay for peroxisomal acyl-CoA oxidase. Biochem J. 1985 Apr 1;227(1):205–210. doi: 10.1042/bj2270205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Spydevold O., Bremer J. Induction of peroxisomal beta-oxidation in 7800 C1 Morris hepatoma cells in steady state by fatty acids and fatty acid analogues. Biochim Biophys Acta. 1989 May 15;1003(1):72–79. doi: 10.1016/0005-2760(89)90101-x. [DOI] [PubMed] [Google Scholar]
  27. Sørensen H. N., Gautik K. M., Bremer J., Spydevold O. Induction of the three peroxisomal beta-oxidation enzymes is synergistically regulated by dexamethasone and fatty acids, and counteracted by insulin in Morris 7800C1 hepatoma cells in culture. Eur J Biochem. 1992 Sep 15;208(3):705–711. doi: 10.1111/j.1432-1033.1992.tb17238.x. [DOI] [PubMed] [Google Scholar]
  28. Sørensen H. N., Hvattum E., Paulssen E. J., Gautvik K. M., Bremer J., Spydevold O. Induction of peroxisomal acyl-CoA oxidase by 3-thia fatty acid, in hepatoma cells and hepatocytes in culture is modified by dexamethasone and insulin. Biochim Biophys Acta. 1993 Jan 23;1171(3):263–271. doi: 10.1016/0167-4781(93)90064-k. [DOI] [PubMed] [Google Scholar]
  29. Topping D. L., Mayes P. A. The immediate effects of insulin and fructose on the metabolism of the perfused liver. Changes in lipoprotein secretion, fatty acid oxidation and esterification, lipogenesis and carbohydrate metabolism. Biochem J. 1972 Jan;126(2):295–311. doi: 10.1042/bj1260295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wang S. R., Pessah M., Infante J., Catala D., Salvat C., Infante R. Lipid and lipoprotein metabolism in Hep G2 cells. Biochim Biophys Acta. 1988 Aug 12;961(3):351–363. doi: 10.1016/0005-2760(88)90082-3. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Wong S., Reardon M., Nestel P. Reduced triglyceride formation from long-chain polyenoic fatty acids in rat hepatocytes. Metabolism. 1985 Oct;34(10):900–905. doi: 10.1016/0026-0495(85)90135-0. [DOI] [PubMed] [Google Scholar]

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