Pretreatment of Macrophages with Paclitaxel Inhibits iNOS Expression

  • Li Mei-Hong (Department of Pharmacology, Chosun University College of Medicine) ;
  • Kang Jong-Soon (Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim Hwan-Mook (Korea Research Institute of Bioscience and Biotechnology) ;
  • Jeon Young-Jin (Department of Pharmacology, Chosun University College of Medicine)
  • Published : 2006.06.01

Abstract

We demonstrate that paclitaxel, an antitumor agent derived from yew tree, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Previously, paclitaxel has been known to induce iNOS gene expression in macrophages. However, in this report we described that the pre-treatment of macrophages with paclitaxel ($0.1{\mu}M$) for 8 h inhibited LPS-induced iNOS gene expression. Pretreatment of RAW 264.7 cells with paclitaxel significantly inhibited LPS-stimulated nitric oxide (NO) production. Western immunoblot of iNOS and RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression in RAW 264.7 cells. Immunocytochemical staining of iNOS further confirmed that pretreatment of macrophages with paclitaxel inhibited macrophage activation. Electrophoretic mobility shift assay showed that paclitaxel inhibited $NF-_{\kappa}/Rel$ DNA binding. Collectively, these series of experiments indicate that paclitaxel inhibits iNOS gene expression by blocking $NF-_{\kappa}B/Rel$ activation.

References

  1. Blagosklonny, M.V., Schulte, T., Nguyen, P., Trepel, J. and Neckers, L.M. (1996): Taxol-induced apoptosis and phosphorylation of Bcl-2 protein involved c-Raf-1 and represents a novel c-Raf-1 signal transduction pathway. Cancer Res., 56, 1851-1854
  2. Chomczynski, P. and Mackey, K. (1995): Substitution of chloroform by bromo-chloropropane in the single-step method of RNA isolation. Anal. Biochem., 225, 163-164 https://doi.org/10.1006/abio.1995.1126
  3. Cvetkovic, I., Miljkovic, D., Vuckovic, O., Harhaji, L., Nikolic, Z., Trajkovic, V. and Mostarica Stojkovic, M. (2004): Taxol activates inducible nitric oxide synthase in rat astrocytes: the role of MAP kinases and NF-$\kappa$B. Cell. Mol. Life Sci.,61, 1167-1175 https://doi.org/10.1007/s00018-004-3408-5
  4. Das, K.C. and White, C.W. (1997): Activation of NF-$\kappa$B by antineoplastic agents. J. Biol. Chem., 272, 14914-14920 https://doi.org/10.1074/jbc.272.23.14914
  5. Ding, A.H., Porteu, F., Sanchez, E. and Nathan, C.F. (1990): Shared actions of endotoxin and taxol on TNF receptors and TNF release Science, 248, 370-372 https://doi.org/10.1126/science.1970196
  6. Dunnett, M. (1955): A multiple comparison procedure for comparing several treatments with a control. J. Am. Statistics, Assoc., 50, 1096-1121 https://doi.org/10.2307/2281208
  7. Green, L.C., Wagner, D.A., Glogowski, J., Skipper, P.L., Wishnok, J.S. and Tannenbaum, S.R. (1982): Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Ana.l Biochem., 126, 131-138 https://doi.org/10.1016/0003-2697(82)90118-X
  8. Hibbs, J.B. Jr., Taintor, R.R. and Vavrin, Z. (1987): Macrophage cytotoxicity: role for L-arginine deiminase and imino nitrogen oxidation to nitrite. Science, 235, 473-476 https://doi.org/10.1126/science.2432665
  9. Kim, Y.M. and Paik, S. (2005): Induction of expression of inducible nitric oxide synthase by taxol in murine macrophage cells. Biochem. Biophys. Res. Commun., 326, 410-416 https://doi.org/10.1016/j.bbrc.2004.11.043
  10. Lowenstein, C.J., Alley, E.W., Raval, P., Snowman, A.M., Snyder, S.H., Russell, S.W. and Murphy, W.J. (1993): Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc. Natl. Acad. Sci. USA, 90, 9730- 9734
  11. Manthey, C.L., Qureshi, N., Stutz, P.L. and Vogel, S.N. (1993): Lipopolysaccharide antagonists block taxol-induced signaling in murine macrophages. J. Exp. Med., 178, 695- 702 https://doi.org/10.1084/jem.178.2.695
  12. Palmer, R.M., Ashton, D.S. and Moncada, S. (1988): Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature, 333, 664-666 https://doi.org/10.1038/333664a0
  13. Perera, P.Y., Qureshi, N. and Vogel, S.N. (1996): Paclitaxel (Taxol)-induced NF-$\kappa$B translocation in murine macrophages. Infect. Immun., 64, 878-884
  14. Rietschel, E.T. and Brade, H. (1992): Bacterial endotoxins. Sci. Am., 267, 54-61 https://doi.org/10.1038/scientificamerican0892-54
  15. Rowinsky, E.K. (1994): Update on the antitumor activity of paclitaxel in clinical trials. Ann. Pharmacother., 28, S18- S22 https://doi.org/10.1177/10600280940280S505
  16. Xie, Q.W., Kashiwabara, Y. and Nathan, C. (1994): Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J. Biol. Chem., 269, 4705-4708