Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Jan 3;92(1):107–111. doi: 10.1073/pnas.92.1.107

Sequential induction of 5-lipoxygenase gene expression and activity in Mono Mac 6 cells by transforming growth factor beta and 1,25-dihydroxyvitamin D3.

M Brungs 1, O Rådmark 1, B Samuelsson 1, D Steinhilber 1
PMCID: PMC42826  PMID: 7816797

Abstract

5-Lipoxygenase (5-LO; EC 1.13.11.34) activity in the human monocytic cell line Mono Mac 6 was upregulated by combined treatment with transforming growth factor beta 1 (TGF-beta) and 1,25-dihydroxyvitamin D3 (VD3). In undifferentiated cells, 5-LO enzyme activity was undetectable. After the addition of TGF-beta plus VD3, the activity of intact cells was 800 ng per 10(6) cells--500 times more than the assay detection limit. Also 5-LO protein and mRNA expression were induced > 128-fold and 64-fold, respectively, as compared to undifferentiated cells. Both TGF-beta and VD3 were required for these prominent responses. Either agent alone gave small amounts of 5-LO protein and mRNA but very low 5-LO activities. After the addition of TGF-beta and VD3, the induction of 5-LO protein was obvious after 1 day, but the increase in activity was delayed and did not appear until the second day. Pretreatment of cells with TGF-beta or VD3 alone for 2 days led to 5-LO protein expression but very low enzyme activity. Addition of the lacking second inducer was required for full induction of 5-LO protein expression and for upregulation of enzyme activity. Partial purification of 5-LO from Mono Mac 6 cells and recombination with soluble cellular proteins from different sources indicated the presence of cytosolic factors that affect the activity of 5-LO.

Full text

PDF
107

Images in this article

108Fig. 1
on p.108
109Fig. 2
on p.109
109Fig. 3
on p.109
110Fig. 5
on p.110

Selected References

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

  1. Bennett C. F., Chiang M. Y., Monia B. P., Crooke S. T. Regulation of 5-lipoxygenase and 5-lipoxygenase-activating protein expression in HL-60 cells. Biochem J. 1993 Jan 1;289(Pt 1):33–39. doi: 10.1042/bj2890033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bigby T. D., Holtzman M. J. Enhanced 5-lipoxygenase activity in lung macrophages compared to monocytes from normal subjects. J Immunol. 1987 Mar 1;138(5):1546–1550. [PubMed] [Google Scholar]
  3. Bonewald L. F., Kester M. B., Schwartz Z., Swain L. D., Khare A., Johnson T. L., Leach R. J., Boyan B. D. Effects of combining transforming growth factor beta and 1,25-dihydroxyvitamin D3 on differentiation of a human osteosarcoma (MG-63). J Biol Chem. 1992 May 5;267(13):8943–8949. [PubMed] [Google Scholar]
  4. Bonser R. W., Siegel M. I., Chung S. M., McConnell R. T., Cuatrecasas P. Esterification of an endogenously synthesized lipoxygenase product into granulocyte cellular lipids. Biochemistry. 1981 Sep 1;20(18):5297–5301. doi: 10.1021/bi00521a032. [DOI] [PubMed] [Google Scholar]
  5. Claesson H. E., Jakobsson P. J., Steinhilber D., Odlander B., Samuelsson B. Expression of 5-lipoxygenase and biosynthesis of leukotriene B4 in human monomorphonuclear leukocytes. J Lipid Mediat. 1993 Mar-Apr;6(1-3):15–22. [PubMed] [Google Scholar]
  6. Coffey M. J., Gyetko M., Peters-Golden M. 1,25-Dihydroxyvitamin D3 upregulates 5-lipoxygenase metabolism and 5-lipoxygenase activating protein in peripheral blood monocytes as they differentiate into mature macrophages. J Lipid Mediat. 1993 Mar-Apr;6(1-3):43–51. [PubMed] [Google Scholar]
  7. Goldyne M. E., Burrish G. F., Poubelle P., Borgeat P. Arachidonic acid metabolism among human mononuclear leukocytes. Lipoxygenase-related pathways. J Biol Chem. 1984 Jul 25;259(14):8815–8819. [PubMed] [Google Scholar]
  8. Kargman S., Rouzer C. A. Studies on the regulation, biosynthesis, and activation of 5-lipoxygenase in differentiated HL60 cells. J Biol Chem. 1989 Aug 5;264(22):13313–13320. [PubMed] [Google Scholar]
  9. Matsumoto T., Funk C. D., Rådmark O., Hög J. O., Jörnvall H., Samuelsson B. Molecular cloning and amino acid sequence of human 5-lipoxygenase. Proc Natl Acad Sci U S A. 1988 Jan;85(1):26–30. doi: 10.1073/pnas.85.1.26. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Morikawa M., Harada N., Soma G., Yoshida T. Transforming growth factor-beta 1 modulates the effect of 1 alpha, 25-dihydroxyvitamin D3 on leukemic cells. In Vitro Cell Dev Biol. 1990 Jul;26(7):682–690. doi: 10.1007/BF02624424. [DOI] [PubMed] [Google Scholar]
  11. Pueringer R. J., Bahns C. C., Hunninghake G. W. Alveolar macrophages have greater amounts of the enzyme 5-lipoxygenase than do monocytes. J Appl Physiol (1985) 1992 Aug;73(2):781–786. doi: 10.1152/jappl.1992.73.2.781. [DOI] [PubMed] [Google Scholar]
  12. Reid G. K., Kargman S., Vickers P. J., Mancini J. A., Léveillé C., Ethier D., Miller D. K., Gillard J. W., Dixon R. A., Evans J. F. Correlation between expression of 5-lipoxygenase-activating protein, 5-lipoxygenase, and cellular leukotriene synthesis. J Biol Chem. 1990 Nov 15;265(32):19818–19823. [PubMed] [Google Scholar]
  13. Samuelsson B., Dahlén S. E., Lindgren J. A., Rouzer C. A., Serhan C. N. Leukotrienes and lipoxins: structures, biosynthesis, and biological effects. Science. 1987 Sep 4;237(4819):1171–1176. doi: 10.1126/science.2820055. [DOI] [PubMed] [Google Scholar]
  14. Steinhilber D., Herrmann T., Roth H. J. Separation of lipoxins and leukotrienes from human granulocytes by high-performance liquid chromatography with a Radial-Pak cartridge after extraction with an octadecyl reversed-phase column. J Chromatogr. 1989 Sep 1;493(2):361–366. doi: 10.1016/s0378-4347(00)82742-5. [DOI] [PubMed] [Google Scholar]
  15. Steinhilber D., Hoshiko S., Grunewald J., Rådmark O., Samuelsson B. Serum factors regulate 5-lipoxygenase activity in maturating HL60 cells. Biochim Biophys Acta. 1993 Jul 28;1178(1):1–8. doi: 10.1016/0167-4889(93)90104-w. [DOI] [PubMed] [Google Scholar]
  16. Steinhilber D., Rådmark O., Samuelsson B. Transforming growth factor beta upregulates 5-lipoxygenase activity during myeloid cell maturation. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5984–5988. doi: 10.1073/pnas.90.13.5984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Testa U., Masciulli R., Tritarelli E., Pustorino R., Mariani G., Martucci R., Barberi T., Camagna A., Valtieri M., Peschle C. Transforming growth factor-beta potentiates vitamin D3-induced terminal monocytic differentiation of human leukemic cell lines. J Immunol. 1993 Mar 15;150(6):2418–2430. [PubMed] [Google Scholar]
  18. Wiseman J. S. Alpha-secondary isotope effects in the lipoxygenase reaction. Biochemistry. 1989 Mar 7;28(5):2106–2111. doi: 10.1021/bi00431a021. [DOI] [PubMed] [Google Scholar]
  19. Zhang Y. Y., Lind B., Rådmark O., Samuelsson B. Iron content of human 5-lipoxygenase, effects of mutations regarding conserved histidine residues. J Biol Chem. 1993 Feb 5;268(4):2535–2541. [PubMed] [Google Scholar]
  20. Ziegler-Heitbrock H. W., Thiel E., Fütterer A., Herzog V., Wirtz A., Riethmüller G. Establishment of a human cell line (Mono Mac 6) with characteristics of mature monocytes. Int J Cancer. 1988 Mar 15;41(3):456–461. doi: 10.1002/ijc.2910410324. [DOI] [PubMed] [Google Scholar]
  21. van den Eijnden-van Raaij A. J., Koornneef I., van Zoelen E. J. A new method for high yield purification of type beta transforming growth factor from human platelets. Biochem Biophys Res Commun. 1988 Nov 30;157(1):16–23. doi: 10.1016/s0006-291x(88)80004-4. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES