Highlights
- •An in vitro model for melanocyte senescence using UVB exposure is proposed.
- •The protocol relies on UVB exposure twice and subsequent 2-week cultivation.
- •Melanocyte senescence accompanies hyperpigmentation via prolonged p53 expression.
Abstract
Background
Ultraviolet radiation (UVR) is a well-known factor in skin aging and pigmentation,
and daily exposure to subcytotoxic doses of UVR might accelerate senescence and senescence-associated
phenomena in human melanocytes.
Objective
To establish an in vitro melanocyte model to mimic the conditions of repeated exposure to subcytotoxic doses
of UVB irradiation and to investigate key factor(s) for melanocyte senescence and
senescence-associated phenomena.
Methods
Human epidermal melanocytes were exposed twice with 20 mJ/cm2 UVB over a 24-h interval and subsequently cultivated for 2 weeks. Senescent phenotypes
were addressed morphologically, and by measuring the senescence-associated β-galactosidase
(SA-β-Gal) activity, cell proliferation capacity with cell cycle analysis, and melanin
content.
Results
The established protocol successfully induced melanocyte senescence, and senescent
melanocytes accompanied hyperpigmentation. Prolonged expression of p53 was responsible
for melanocyte senescence and hyperpigmentation, and treatment with the p53-inhibitor
pifithrin-α at 2-weeks post-UVB irradiation, but not at 48 h, significantly reduced
melanin content along with decreases in tyrosinase levels.
Conclusion
Melanocyte senescence model will be useful for studying the long-term effects of UVB
irradiation and pigmentation relevant to physiological photoaging, and screening compounds
effective for senescence-associated p53-mediated pigmentation.
Abbreviations:
UVR (ultraviolet radiation), UVsen (UVB-induced senescence), SAP (senescence-associated pigmentation), SA-β-Gal (senescence-associated β-galactosidase), TPA (tetradecanoyl phorbol acetate), CT (cholera toxin), pTpT (thymine dinucleotides), HEMn (human epidermal melanocyte neonatal), CDKN1A (cyclin dependent kinase inhibitor 1A), PFTα (pifithrin-α), TYR (tyrosinase), TYRP1 (tyrosinase-related protein 1), POMC (pro-opiomelanocortin), α-MSH (α-melanocyte-stimulating hormone), KITLG (c-KIT ligand), HGF (hepatocyte growth factor)Keywords
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Article info
Publication history
Published online: February 26, 2018
Accepted:
February 22,
2018
Received in revised form:
December 29,
2017
Received:
September 15,
2017
Identification
Copyright
© 2018 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.
