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Hmgb1 Promotes Wound Healing of 3T3 Mouse Fibroblasts via Rage-Dependent ERK1/2 Activation

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Abstract

HMGb1 is a nuclear protein playing a role in DNA architecture and transcription. This protein has also been shown to function as a cytokine and to stimulate keratinocyte scratch wound healing. Due to the importance of finding new wound healing molecules, we have studied the effects of HMGb1 on fibroblasts, another major skin cell type, using the NIH 3T3 line. HMGb1 expression in these cells was assessed by Western blot, while its nuclear localization was pointed out by confocal immunofluorescence. HMGb1-induced cell proliferation with a maximum at a concentration of 10 nM, and such a dose also stimulated cell migration and scratch wound healing. Western blot analysis showed that HMGb1 activates ERK1/2, while the use of an anti-RAGE receptor-blocking antibody and of the selective MEK1/2 inhibitor PD98059 blocked ERK1/2 activation and wound healing responses to HMGb1. Taken together data show that HMGb1 promotes 3T3 fibroblast wound healing by inducing cell proliferation and migration, and that this occurs through the activation of the RAGE/MEK/ERK pathway. In conclusion, HMGb1 seems a good candidate for the development of medical treatments to be used on chronic or severe wounds.

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References

  1. Goodwin, G., Sanders, C., & Johns, E. (1973). A new group of chromatin-associated proteins with a high content of acidic and basic amino acids. European Journal of Biochemistry, 38, 14–19.

    Article  PubMed  CAS  Google Scholar 

  2. Li, J., Kokkola, R., Tabibzadeh, S., Yang, R., Ochani, M., Qiang, X., et al. (2003). Structural basis for the proinflammatory cytokine activity of high mobility group box 1. Molecular Medicine, 9, 37–45.

    PubMed  CAS  Google Scholar 

  3. Wang, H., Bloom, O., Zhang, M., Vishnubhakat, J. M., Ombrellino, M., Che, J., et al. (1999). HMG-1 as a late mediator of endotoxin lethality in mice. Science, 285, 248–251.

    Article  PubMed  CAS  Google Scholar 

  4. Scaffidi, P., Misteli, T., & Bianchi, M. E. (2002). Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature, 418, 191–195.

    Article  PubMed  CAS  Google Scholar 

  5. Lotze, M. T., & Tracey, K. J. (2005). High-mobility group box 1 protein (HMGB1): Nuclear weapon in the immune arsenal. Nature Reviews Immunology, 5, 331–342.

    Article  PubMed  CAS  Google Scholar 

  6. Degryse, B., Bonaldi, T., Scaffidi, P., Muller, S., Resnati, M., Sanvito, F., et al. (2001). The high mobility group (HMG) boxes of the nuclear protein HMG1 induce chemotaxis and cytoskeleton reorganization in rat smooth muscle cells. Journal of Cell Biology, 152, 1197–1206.

    Article  PubMed  CAS  Google Scholar 

  7. Palumbo, R., Sampaolesi, M., De Marchis, F., Tonlorenzi, R., Colombetti, S., Mondino, A., et al. (2004). Extracellular HMGB1, a signal of tissue damage, induces mesoangioblast migration and proliferation. Journal of Cell Biology, 164, 441–449.

    Article  PubMed  CAS  Google Scholar 

  8. Chavakis, E., Hain, A., Vinci, M., Carmona, G., Bianchi, M. E., Vajkoczy, P., et al. (2007). High-mobility group box 1 activates integrin-dependent homing of endothelial progenitor cells. Circulation Research, 100, 204–212.

    Article  PubMed  CAS  Google Scholar 

  9. Rauvala, H., & Rouhiainen, A. (2007). RAGE as a receptor of HMGB1 (Amphoterin): Roles in health and disease. Current Molecular Medicine, 7, 725–734.

    Article  PubMed  CAS  Google Scholar 

  10. Martin, P. (1997). Wound healing–aiming for perfect skin regeneration. Science, 276, 75–81.

    Article  PubMed  CAS  Google Scholar 

  11. Welt, K., Hinrichs, R., Weiss, J., Burgdorf, W., Krieg, T., & Scharffetter-Kochanek, K. (2009). Wound healing. European Journal of Dermatology, 19, 413–416.

    PubMed  CAS  Google Scholar 

  12. Declair, V. (1999). The importance of growth factors in wound healing. Ostomy Wound Management, 45, 64–74.

    CAS  Google Scholar 

  13. Broughton, G., Janis, J. E., & Attinger, C. E. (2006). The basic science of wound healing. Plastic and Reconstructive Surgery, 117, 12–34.

    Article  CAS  Google Scholar 

  14. Ranzato, E., Patrone, M., Pedrazzi, M., & Burlando, B. (2009). HMGb1 promotes scratch wound closure of HaCaT keratinocytes via ERK1/2 activation. Molecular and Cellular Biochemistry, 332, 199–205.

    Article  PubMed  CAS  Google Scholar 

  15. Todaro, G. J., & Green, H. (1963). Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. Journal of Cell Biology, 17, 299–313.

    Article  PubMed  CAS  Google Scholar 

  16. DeBiasio, R., Bright, G. R., Ernst, L. A., Waggoner, A. S., & Taylor, D. L. (1987). Five-parameter fluorescence imaging: Wound healing of living Swiss 3T3 cells. Journal of Cell Biology, 105, 1613–1622.

    Article  PubMed  CAS  Google Scholar 

  17. Bassi, R., Giussani, P., Anelli, V., Colleoni, T., Pedrazzi, M., Patrone, M., et al. (2008). HMGB1 as an autocrine stimulus in human T98G glioblastoma cells: Role in cell growth and migration. Journal of Neuro-oncology, 87, 23–33.

    Article  PubMed  CAS  Google Scholar 

  18. Ishihara, K., Tsutsumi, K., Kawane, S., Nakajima, M., & Kasaoka, T. (2003). The receptor for advanced glycation end-products (RAGE) directly binds to ERK by a D-domain-like docking site. FEBS Letters, 550, 107–113.

    Article  PubMed  CAS  Google Scholar 

  19. Kokkola, R., Andersson, A., Mullins, G., Ostberg, T., Treutiger, C. J., Arnold, B., et al. (2005). RAGE is the major receptor for the proinflammatory activity of HMGB1 in rodent macrophages. Scandinavian Journal of Immunology, 61, 1–9.

    Article  PubMed  CAS  Google Scholar 

  20. Sparatore, B., Passalacqua, M., Patrone, M., Melloni, E., & Pontremoli, S. (1996). Extracellular high-mobility group 1 protein is essential for murine erythroleukaemia cell differentiation. Biochemical Journal, 320, 253–256.

    PubMed  CAS  Google Scholar 

  21. Pedrazzi, M., Patrone, M., Passalacqua, M., Ranzato, E., Colamassaro, D., Sparatore, B., et al. (2007). Selective proinflammatory activation of astrocytes by high-mobility group box 1 protein signaling. Journal of Immunology, 179, 8525–8532.

    CAS  Google Scholar 

  22. Sparatore, B., Patrone, M., Passalacqua, M., Pedrazzi, M., Ledda, S., Pontremoli, S., et al. (2005). Activation of A431 human carcinoma cell motility by extracellular high-mobility group box 1 protein and epidermal growth factor stimuli. Biochemical Journal, 389, 215–221.

    Article  PubMed  CAS  Google Scholar 

  23. Borenfreund, E., Babich, H., & Martin-Alguacil, N. (1988). Comparison of two in vitro cytotoxicity assays: The neutral red (NR) and tetrozolium (MTT) tests. Toxicology in Vitro, 2, 1–6.

    Article  CAS  Google Scholar 

  24. Ekunwe, S. I., Hunter, R. D., & Hwang, H. M. (2005). Ultraviolet radiation increases the toxicity of pyrene, 1-aminopyrene and 1-hydroxypyrene to human keratinocytes. Internatioanl Journal of Environmental Research and Public Health, 2, 58–62.

    Article  CAS  Google Scholar 

  25. Ellerman, J. E., Brown, C. K., de Vera, M., Zeh, H. J., Billiar, T., Rubartelli, A., et al. (2007). Masquerader: High mobility group box-1 and cancer. Clin Cancer Res, 13, 2836–2848.

    Article  PubMed  CAS  Google Scholar 

  26. Urbonaviciute, V., Furnrohr, B. G., Weber, C., Haslbeck, M., Wilhelm, S., Herrmann, M., et al. (2007). Factors masking HMGB1 in human serum and plasma. Journal of Leukocyte Biology, 81, 67–74.

    Article  PubMed  CAS  Google Scholar 

  27. Zimmermann, K., Volkel, D., Pable, S., Lindner, T., Kramberger, F., Bahrami, S., et al. (2004). Native versus recombinant high-mobility group B1 proteins: Functional activity in vitro. Inflammation, 28, 221–229.

    Article  PubMed  CAS  Google Scholar 

  28. Hamada, N., Maeyama, T., Kawaguchi, T., Yoshimi, M., Fukumoto, J., Yamada, M., et al. (2008). The role of high mobility group box1 in pulmonary fibrosis. American Journal of Respiratory Cell and Molecular Biology, 39, 440–447.

    Article  PubMed  CAS  Google Scholar 

  29. Straino, S., Di Carlo, A., Mangoni, A., De Mori, R., Guerra, L., Maurelli, R., et al. (2008). High-mobility group box 1 protein in human and murine skin: Involvement in wound healing. Journal of Investigative Dermatology, 128, 1545–1553.

    Article  PubMed  CAS  Google Scholar 

  30. Porto, A., Palumbo, R., Pieroni, M., Aprigliano, G., Chiesa, R., Sanvito, F., et al. (2006). Smooth muscle cells in human atherosclerotic plaques secrete and proliferate in response to high mobility group box 1 protein. The FASEB Journal, 20, 2565–2566.

    Article  PubMed  CAS  Google Scholar 

  31. Yang, D., Chen, Q., Yang, H., Tracey, K. J., Bustin, M., & Oppenheim, J. J. (2007). High mobility group box-1 protein induces the migration and activation of human dendritic cells and acts as an alarmin. Journal of Leukocyte Biology, 81, 59–66.

    Article  PubMed  CAS  Google Scholar 

  32. Yu, M., Wang, H., Ding, A., Golenbock, D. T., Latz, E., Czura, C. J., et al. (2006). HMGB1 signals through toll-like receptor (TLR) 4 and TLR2. Shock, 26, 174–179.

    Article  PubMed  CAS  Google Scholar 

  33. Eming, S. A., Krieg, T., & Davidson, J. M. (2007). Inflammation in wound repair: Molecular and cellular mechanisms. Journal of Investigative Dermatology, 127, 514–525.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by a grant from Ricerca Sanitaria Finalizzata, Regione Piemonte, Italy, 2008 bis, and by grants from the University of Piemonte Orientale “Amedeo Avogadro”. ER is recipient of a Research Fellowship from the University of Piemonte Orientale.

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Authors and Affiliations

  1. Department of Environment and Life Sciences, DiSAV, University of Piemonte Orientale, Viale T. Michel 11, 15121, Alessandria, Italy

    Elia Ranzato, Mauro Patrone & Bruno Burlando

  2. Molecular Histology and Cell Growth Unit, San Raffaele Scientific Institute, Via Olgettina 58, 20123, Milan, Italy

    Elia Ranzato

  3. Department of Experimental Medicine-Biochemistry Section and Centre of Excellence for Biomedical Research, University of Genoa, Viale Benedetto XV 1, 16132, Genoa, Italy

    Marco Pedrazzi

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  1. Elia Ranzato
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  2. Mauro Patrone
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  4. Bruno Burlando
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Correspondence to Elia Ranzato.

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Ranzato, E., Patrone, M., Pedrazzi, M. et al. Hmgb1 Promotes Wound Healing of 3T3 Mouse Fibroblasts via Rage-Dependent ERK1/2 Activation. Cell Biochem Biophys 57, 9–17 (2010). https://doi.org/10.1007/s12013-010-9077-0

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  • DOI: https://doi.org/10.1007/s12013-010-9077-0

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