Skip to main content
SpringerLink
Log in

Effects of n−3 and n−6 fatty acids on tumor necrosis factor cytotoxicity in WEHI fibrosarcoma cells

  • Article
  • Published:
Lipids

Abstract

Modulation by fatty acids of the cytotoxic effect of recombinant tumor necrosis factor alpha (TNF) toward WEHI 164 mouse fibrosarcoma cells has been examined. Preincubating the highly TNF-sensitive WEHI clone 13 cells for 44 hr with 50 μmol/L of 20∶5n−3, 22∶6n−3, 18∶3n−6, 20∶3n−6 or 20∶4n−6 reduced cell survival 22 hr after challenge with TNF (40 ng/L) by 65%, 72%, 60%, 98% and 85%, respectively. In comparison, 18∶3n−3, 18∶2n−6 and 18∶1n−9 had only negligible effects on TNF-induced toxicity. Different extent of fatty acid incorporation into cell total phospholipids or triglycerides could not explain the observed effects on TNF cytotoxicity, and the enhanced cytotoxicity could therefore not be explained merely by an increased unsaturation of the cell membranes. In addition to the fatty acid supplied, preincubation with 18∶2n−6, 18∶3n−6 or 18∶3n−3 also enriched the cells with 20∶2n−6, 20∶3n−6 and 20∶3n−3, respectively, most likely due to chain elongation. The results suggest that the WEHI cells have a low Δ6 desaturase activity, and that n−6 and n−3 acids must have at least 3 or 4 double bonds, respectively, to enhance TNF cytotoxicity in WEHI cells. Dexamethasone partly inhibited TNF-induced cytotoxicity, while cyclooxygenase, thromboxane synthetase or lipoxygenase inhibitors had no or negligible effects. The antioxidant butylated hydroxyanisole (BHA) completely inhibited TNF-induced cytotoxicity, while the structurally and functionally similar antioxidant butylated hydroxy-toluene had no such effect, indicating that BHA does not block TNF cytotoxicity through its antioxidant effect. The results suggest that TNF cytotoxicity involves, directly or indirectly, metabolism of long-chain polyun-saturated fatty acids, and we speculate that fatty acid metabolites are involved.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BHA:

butylated hydroxyanisole

BHT:

butylated hydroxytoluene

ED50 :

defined as the TNF concentration that reduces cell density by 50%

FAME:

fatty acid methyl ester(s)

FCS-M:

RPMI-1640 medium containing 10% (v/v) heat-inactivated fetal calf serum, 2 mmol/L L-glutamine and 40 mg/L genetamicin

GLC:

gas-liquid chromatography

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NDGA:

nordihydroguaretic acid

SDS:

sodium dodecyl sulfate

TLC:

thin-layer chromatography

TNF:

recombinant murine tumor necrosis factor alpha

References

  1. Le, J., and Vilček, J. (1987)Lab. Invest. 56 234–248.

    PubMed  CAS  Google Scholar 

  2. Nissen-Meyer, J., Austgulen, R., and Espevik, T. (1987)Cancer Res. 47, 2251–2258.

    PubMed  CAS  Google Scholar 

  3. Sugarman, B.J., Aggarwal, B.B., Hass, P.E., Figari, I.S., Palladino, Jr., M.A., and Shepard, H.M. (1985)Science 230, 943–945.

    Article  PubMed  CAS  Google Scholar 

  4. Urban, J.L., Shepard, H.M., Rothstein, J.L., Sugarman, B.J., and Schreiber, H. (1986)Proc. Natl. Acad. Sci. USA 83, 5233–5237.

    Article  PubMed  CAS  Google Scholar 

  5. Tsujimoto, M., Yip, Y.K., and Vilček, J. (1985)Proc. Natl. Acad. Sci. USA 82, 7626–7630.

    Article  PubMed  CAS  Google Scholar 

  6. Johnson, S.E., and Baglioni, C. (1988)J. Biol. Chem. 263, 5686–5692.

    PubMed  CAS  Google Scholar 

  7. Creasy, A.A., Yamamoto, R., and Vitt, C.R. (1987)Proc. Natl. Acad. Sci. USA 84, 3293–3297.

    Article  Google Scholar 

  8. Zimmermann, R.J., Marafino, B.J., Chan, A., Landre, P., and Winkelhake, J.L. (1989)J. Immunol. 142, 1405–1409.

    Google Scholar 

  9. Kull, F.C. (1988)Biochem. Biophys. Res. Commun. 153, 402–409.

    Article  PubMed  CAS  Google Scholar 

  10. Hepburn, A., Boeynaems, J.M., Fiers, W., and Dumont, J.E. (1987)Biochem. Biophys. Res. Commun. 149, 815–822.

    Article  PubMed  CAS  Google Scholar 

  11. Suffys, P., Beyaert, R., Van Roy, F., and Fiers, W. (1987)Biochem. Biophys. Res. Commun. 149, 735–743.

    Article  PubMed  CAS  Google Scholar 

  12. Nielsen, N.H., and Hansen, J.P.H. (1980)J. Cancer. Res. Clin. Oncol. 98, 287–299.

    Article  PubMed  CAS  Google Scholar 

  13. Carroll, K.K., and Braden, L.M. (1986)Prog. Lipid Res. 25, 583–585.

    Article  CAS  Google Scholar 

  14. Welsch, C.W. (1987)Am. J. Clin. Nutr. 45, 192–202.

    PubMed  CAS  Google Scholar 

  15. Schlager, S.I., and Ohanian, S.H. (1979)J. Immunol. 123, 146–152.

    PubMed  CAS  Google Scholar 

  16. Espevik, T., and Nissen-Meyer, J. (1986)J. Immunol. Methods 95, 99–105.

    Article  PubMed  CAS  Google Scholar 

  17. Metcalf, L.D., and Schmitz, A.A. (1961)Anal. Chem. 33, 363–364.

    Article  Google Scholar 

  18. Mosmann, T. (1983)J. Immunol. Methods 65, 55–63.

    Article  PubMed  CAS  Google Scholar 

  19. Brunner, K.T., Mauel, J., Cerottini, J.C., and Chapuis, B. (1968)Immunology 14, 181–196.

    PubMed  CAS  Google Scholar 

  20. Kaluzny, M.A., Duncan, L.A., Merritt, M.V., and Epps, D.E. (1985)J. Lipid Res. 26, 135–140.

    PubMed  CAS  Google Scholar 

  21. Bjerve, K.S., Fischer, S., and Alme, K. (1987)Am. J. Clin. Nutr. 46, 570–576.

    PubMed  CAS  Google Scholar 

  22. Bligh, E.G., and Dyer, W.J. (1959)Can. J. Biochem. Physiol. 37, 911–917.

    PubMed  CAS  Google Scholar 

  23. Mangold, H.K. (1962) inDünnschichtchromatographie (Stahl, E., ed.) pp. 156–158, Springer-Verlag, Berlin.

    Google Scholar 

  24. Yamauchi, N., Kuriyama, H., Watanabe, N., Neda, H., Maeda, M., and Niitsu, Y. (1989)Cancer Res. 49, 1671–1675.

    PubMed  CAS  Google Scholar 

  25. Bernert, Jr., J.T., and Sprecher, H. (1975)Biochim. Biophys. Acta 398, 354–363.

    PubMed  Google Scholar 

  26. Reid, T., Ramesha, C.S., and Ringold, G.M. (1991)J. Biol. Chem. 266, 16580–16586.

    PubMed  CAS  Google Scholar 

  27. Das, U.N., Begin, M.E., Ells, G., Huang, Y.S., and Horrobin, D.F. (1987)Biochem. Biophys. Res. Commun. 145, 15–24.

    Article  PubMed  CAS  Google Scholar 

  28. Begin, M.E., Das, U.N., Ells, G., and Horrobin, D.F. (1985)Prostagl. Leukotr. Med. 19, 177–186.

    Article  CAS  Google Scholar 

  29. Chow, S.C., Sisfontes, L., Björkhem, I., and Jondal, M. (1989)Lipids 24, 700–704.

    PubMed  CAS  Google Scholar 

  30. Matthews, N., Neale, M.L., Jackson, S.K., and Stark, J.M. (1987)Immunology 62, 153–155.

    PubMed  CAS  Google Scholar 

  31. Watanabe, N., Niitsu, Y., Neda, H., Sone, H., Yamauchi, N., Maeda, M., and Urushizaki, I. (1988)Immunopharmacol. Immunotoxicol. 10, 109–116.

    PubMed  CAS  Google Scholar 

  32. Schewe, T., Kühn, H., and Rapoport, S.M. (1987) inProstaglandins and Related Substances: A Practical Approach (Benedetto, C., McDonald-Gibson, R.G., Nigam, S. and Slater, T.F., eds.) pp. 229–242, IRL Press, Oxford.

    Google Scholar 

  33. Thompson, D., and Eling, T. (1989)Molec. Pharmacol. 36, 809–817.

    CAS  Google Scholar 

  34. Hori, T., Kashiyama, S., Hayakawa, M., Shibamoto, S., Tsujimoto, M., Oku, N., and Ito, F. (1989)Exp. Cell. Res. 185, 41–49.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Clinical Chemistry, University Hospital, University of Trondheim, N-7006, Trondhelm, Norway

    Ole-L. Brekke, Tora Bardal & Kristian S. Bjerve

  2. Institute of Cancer Research, University Hospital, University of Trondheim, N-7006, Trondhelm, Norway

    Terje Espevik

Authors
  1. Ole-L. Brekke
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Terje Espevik
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tora Bardal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kristian S. Bjerve
    View author publications

    You can also search for this author in PubMed Google Scholar

About this article

Cite this article

Brekke, OL., Espevik, T., Bardal, T. et al. Effects of n−3 and n−6 fatty acids on tumor necrosis factor cytotoxicity in WEHI fibrosarcoma cells. Lipids 27, 161–168 (1992). https://doi.org/10.1007/BF02536172

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02536172

Keywords

Search

Navigation