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HMGb1 promotes scratch wound closure of HaCaT keratinocytes via ERK1/2 activation

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Abstract

HMGb1 is a DNA-binding protein whose role as an extracellular cytokine in inflammation and tissue regeneration has also been reported. Given the importance of keratinocytes in wound healing, we have studied the mechanism of action of HMGb1 on HaCaT keratinocytes during in vitro scratch wound repair. Western blot and confocal immunofluorescence microscopy showed that these cells express significant amounts of HMGb1, that the protein is prevalently localized in the nucleus, and that its release by cells is negligible. Western blot also showed that these cells express the HMGb1 receptor RAGE. Cell exposure to HMGb1 in the absence of serum resulted in a stimulation of cell proliferation and ERK1/2 activation. HMGb1 also accelerated the wound closure of scratch wounded cells and promoted cell migration, as evaluated by a transwell assay. The HMGb1-induced increases of cell proliferation, cell migration, and wound closure were abolished by the MEK inhibitor PD98059. Taken together, data show that, although HMGb1 is not released by HaCaT, when applied exogenously it can induce a marked increase of the wound repair of these cells. Data also suggest that HMGb1 acts via the RAGE/MEK/ERK pathway. These results bring scientific support to the potential application of HMGb1 in regenerative medicine.

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

    Marco Pedrazzi

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  1. Elia Ranzato
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  2. Mauro Patrone
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Correspondence to Elia Ranzato.

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Ranzato, E., Patrone, M., Pedrazzi, M. et al. HMGb1 promotes scratch wound closure of HaCaT keratinocytes via ERK1/2 activation. Mol Cell Biochem 332, 199–205 (2009). https://doi.org/10.1007/s11010-009-0192-4

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  • DOI: https://doi.org/10.1007/s11010-009-0192-4

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