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Activation of RAW264.7 mouse macrophage cells in vitro through treatment with recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB and JAK-STAT kinase signaling pathways

  • Authors:
    • Na Xu
    • Hongyan Yuan
    • Wensen Liu
    • Songyan Li
    • Yang Liu
    • Jiayu Wan
    • Xiaoyan Li
    • Rui Zhang
    • Yaping Chang

  • View Affiliations

  • Published online on: July 2, 2013     https://doi.org/10.3892/ijmm.2013.1426
  • Pages: 729-735
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Abstract

Ricin toxin-binding subunit B (RTB) is a galactose-binding lectin protein. In the present study, we investigated the effects of RTB on inducible nitric oxide (NO) synthase (iNOS), interleukin (IL)-6 and tumor necrosis factor (TNF)-α, as well as the signal transduction mechanisms involved in recombinant RTB-induced macrophage activation. RAW264.7 macrophages were treated with RTB. The results revealed that the mRNA and protein expression of iNOS was increased in the recombinant RTB-treated macrophages. TNF-α production was observed to peak at 20 h, whereas the production of IL-6 peaked at 24 h. In another set of cultures, the cells were co-incubated with RTB and the tyrosine kinase inhibitor, genistein, the phosphatidylinositol 3-kinase (PI3K) inhibitor, LY294002, the p42/44 inhibitor, PD98059, the p38 inhibitor, SB203580, the JNK inhibitor, SP600125, the protein kinase C (PKC) inhibitor, staurosporine, the JAK2 inhibitor, tyrphostin (AG490), or the NOS inhibitor, L-NMMA. The recombinant RTB-induced production of NO, TNF-α and IL-6 was inhibited in the macrophages treated with the pharmacological inhibitors genistein, LY294002, staurosporine, AG490, SB203580 and BAY 11-7082, indicating the possible involvement of protein tyrosine kinases, PI3K, PKC, JAK2, p38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB in the above processes. A phosphoprotein analysis identified tyrosine phosphorylation targets that were uniquely induced by recombinant RTB and inhibited following treatment with genistein; some of these proteins are associated with the downstream cascades of activated JAK-STAT and NF-κB receptors. Our data may help to identify the most important target molecules for the development of novel drug therapies.
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September 2013
Volume 32 Issue 3

Print ISSN: 1107-3756
Online ISSN:1791-244X

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  • Journal Metrics
  • Impact Factor: 5.7
  • Ranked Medicine Research and Experimental
    (total number of cites)
  • CiteScore: 12.3
  • CiteScore Rank: Q1: #28/347 Biochemistry, Genetics and Molecular Biology
  • Source Normalized Impact per Paper (SNIP): 1.022
  • SCImago Journal Rank (SJR): 1.167
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Spandidos Publications style
Xu N, Yuan H, Liu W, Li S, Liu Y, Wan J, Li X, Zhang R and Chang Y: Activation of RAW264.7 mouse macrophage cells in vitro through treatment with recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB and JAK-STAT kinase signaling pathways. Int J Mol Med 32: 729-735, 2013.
APA
Xu, N., Yuan, H., Liu, W., Li, S., Liu, Y., Wan, J. ... Chang, Y. (2013). Activation of RAW264.7 mouse macrophage cells in vitro through treatment with recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB and JAK-STAT kinase signaling pathways. International Journal of Molecular Medicine, 32, 729-735. https://doi.org/10.3892/ijmm.2013.1426
MLA
Xu, N., Yuan, H., Liu, W., Li, S., Liu, Y., Wan, J., Li, X., Zhang, R., Chang, Y."Activation of RAW264.7 mouse macrophage cells in vitro through treatment with recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB and JAK-STAT kinase signaling pathways". International Journal of Molecular Medicine 32.3 (2013): 729-735.
Chicago
Xu, N., Yuan, H., Liu, W., Li, S., Liu, Y., Wan, J., Li, X., Zhang, R., Chang, Y."Activation of RAW264.7 mouse macrophage cells in vitro through treatment with recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB and JAK-STAT kinase signaling pathways". International Journal of Molecular Medicine 32, no. 3 (2013): 729-735. https://doi.org/10.3892/ijmm.2013.1426