In vivo relative quantitative proteomics reveals HMGB1 as a downstream mediator of oestrogen-stimulated keratinocyte migration

Exp Dermatol. 2015 Jun;24(6):478-80. doi: 10.1111/exd.12713. Epub 2015 Apr 27.

Abstract

It is known that oestrogen influences skin wound healing by modulating the inflammatory response, cytokine expression and extracellular matrix deposition; accelerating re-epithelialization; and stimulating angiogenesis. To identify novel proteins associated with effects of oestrogen on keratinocyte, stable isotope labelling by amino acids in cell culture (SILAC)-based mass spectrometry was performed. Using SILAC, quantification of 1085 proteins was achieved. Among these proteins, 60 proteins were upregulated and 32 proteins were downregulated. Among significantly upregulated proteins, high-mobility group protein B1 (HMGB1) has been further evaluated for its role in the effect of oestrogen on keratinocytes. HMGB1 expression was strongly induced in oestrogen-treated keratinocytes in dose- and time-dependent manner. Further, HMGB1 was able to significantly accelerate the rate of HaCaT cell migration. To determine whether HMGB1 is involved in E2-induced HaCaT cell migration, cells were transfected with HMGB1 siRNA. Knockdown of HMGB1 blocked oestrogen-induced keratinocyte migration. Collectively, these experiments demonstrate that HMGB1 is a novel downstream mediator of oestrogen-stimulated keratinocyte migration.

Keywords: HMGB1; estrogen; keratinocyte.

Publication types

  • Letter

MeSH terms

  • Cell Line
  • Cell Movement / drug effects*
  • Cell Movement / physiology
  • Dose-Response Relationship, Drug
  • Estrogens / pharmacology*
  • Gene Expression Regulation / drug effects
  • Gene Expression Regulation / genetics
  • Gene Knockdown Techniques
  • HMGB1 Protein / drug effects
  • HMGB1 Protein / genetics
  • HMGB1 Protein / physiology*
  • Humans
  • Keratinocytes / cytology
  • Keratinocytes / drug effects*
  • Keratinocytes / physiology
  • Proteomics / methods*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / pharmacology
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Time Factors

Substances

  • Estrogens
  • HMGB1 Protein
  • RNA, Small Interfering