The effects of NB-UVB on the hair follicle-derived neural crest stem cells differentiating into melanocyte lineage in vitro

J Dermatol Sci. 2012 Apr;66(1):20-8. doi: 10.1016/j.jdermsci.2012.01.012. Epub 2012 Feb 16.

Abstract

Background: Narrow-band UVB (NB-UVB) is an effective therapeutic option in the treatment of vitiligo. Despite the apparent clinical efficacy, the underlying mechanism of how topical NB-UVB induces repigmentation in vitiligo has not been clearly elucidated.

Objectives: To investigate the effects of NB-UVB on the maturation of melanocyte lineage differentiated from hair follicle-derived neural crest stem cells (HF-NCSCs) in vitro.

Methods: HF-NCSCs were isolated from mouse whisker follicles. The isolated cells were multipotent and expressed embryonic NCSC biomarkers. The effects of NB-UVB on development and differentiation of HF-NCSCs were evaluated. We assessed cell viability, melanogenesis and migration of melanocytes derived from HF-NCSCs after NB-UVB radiation. Tyrosinase, Tyrp1, Dct, Kit, Mc1R, Fzd4, NT3R, Ednra, EP1, TGFβR, Sox10, Mitf, Lef1 and Pax3 gene expression was measured by quantitative RT-PCR, while Tyrosinase, Sox10 and Mitf protein expression were measured by Western blot analysis. Cell migration was measured by Boyden chamber transwell assay.

Results: NB-UVB increased the expression of tyrosinase during melanocytic differentiation from mouse HF-NCSCs, however, NB-UVB inhibited proliferation of melanocytes derived from HF-NCSCs. Mechanistically, increased melanocyte maturation after NB-UVB treatment was resulted from increased expression of several key melanogenic factors, including Sox10, Kit and Mc1R, which play a critical role to promote tyrosinase expression. Furthermore, the migration of the HF-NCSCs-derived melanocytes was downregulated as NB-UVB doses increased. However, the migration of HF-NCSCs was upregulated under 0.4J NB-UVB radiation.

Conclusions: Those data provide in vitro evidence demonstrating some direct effects of NB-UVB on pigmentation of melanocyte lineage differentiated from HF-NCSCs, and may provide a possible mechanism for the effect of NB-UVB in vitiligo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Adhesion / physiology
  • Cell Adhesion / radiation effects
  • Cell Differentiation / physiology
  • Cell Differentiation / radiation effects
  • Cell Lineage / physiology
  • Cell Lineage / radiation effects
  • Cell Movement / physiology
  • Cell Movement / radiation effects
  • Dose-Response Relationship, Radiation
  • Hair Follicle / cytology*
  • Hair Follicle / radiation effects*
  • In Vitro Techniques
  • Intramolecular Oxidoreductases / genetics
  • Intramolecular Oxidoreductases / metabolism
  • Melanocytes / cytology*
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Monophenol Monooxygenase / genetics
  • Monophenol Monooxygenase / metabolism
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / metabolism
  • Neural Stem Cells / radiation effects*
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism
  • Proto-Oncogene Proteins c-kit / genetics
  • Proto-Oncogene Proteins c-kit / metabolism
  • SOXE Transcription Factors / genetics
  • SOXE Transcription Factors / metabolism
  • Transcriptional Activation / physiology
  • Transcriptional Activation / radiation effects
  • Ultraviolet Therapy / methods*

Substances

  • Membrane Glycoproteins
  • SOXE Transcription Factors
  • Sox10 protein, mouse
  • Oxidoreductases
  • Tyrp1 protein, mouse
  • Monophenol Monooxygenase
  • Proto-Oncogene Proteins c-kit
  • Intramolecular Oxidoreductases
  • dopachrome isomerase