130504 paper stduy 한울
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Transcript of 130504 paper stduy 한울
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130504 Park Han-Wool
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Fig. 1. The melanocyte and the tanning response. UVR = ultraviolet radiation,HBD3 = human β-defensin 3, END1 = endothelin 1, SCF = stem cell factor, CAT =catalase, bFGF = basic fibroblast growth factor, MITF = microphthalmia transcriptionfactor, DCT = dopachrome tautomerase, USF-1 = upstream transcription factor 1.
Exposure of human skin to ultraviolet radiation
results in the release of various factors by ker-
atinocytes and melanocytes. Proopiomelanocortin
(POMC) is converted to α-melanocyte stimulating
hormone (α-MSH), which stimulatesmelanocort-
inMC1 receptor (MC1R) cAMP signalling in the
melanocyte causing the upregulation of genes in-
volved in melanogenesis, ultimately resulting in
the tanning response.
Agouti signaling protein (ASIP) antagonises the
action ofα-MSH.Dotted lines and ? indicate inter-
actions that are in need of further research. ROS
= reactive oxygen species.
1. Introduction
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2. Melanocortin MC1 receptor in vitro studies in melanoma andnon-melanocytic cell lines
Fig. 2. cAMP accumulation in B16 or HEK293 cells stably trans-fected with melanocortin MC1 receptor (HEKWTe).
Cells were incubated in serum-free media for at least 2 h. Cells were pre-incubated with 0.1 mM IBMX for 15 min, then stimulated with the indicated ligands for 10 min.
A) 0.5 nM or 1 nM NDP-MSH alone or in combination with 100 nM agouti signaling protein peptide (ASIP-YY) or HBD3, or 100 nM ASIP-YY or HBD3 alone.
B) 1 nM NDP-MSH, 100 nM HBD3 or 100 nM ASIP-YY.
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Fig. 3. ERK phosphorylation in response to α-melanocyte stimulating hormone and human β-Defensin 3.pERK is specific to the phosphorylated Thr202/Tyr204 sites of ERK1/2, ERK recognises all forms of the ERK protein. Anti-GAPDH (R&D Systems) was used as a loading control. Lanes are as indicated C = Control, N = 10 nM NDP-MSH, β =10 nM HBD3.
HEK293 untransfected cells (parental) or HEK293
stably expressing melanocortin MC1 receptor (WTe)
were pre-incubated in serum-free media for at least 2
h, then stimulated with the indicated ligands for 5
min.
Human β-defensin 3 may activate ERK1/2 in HEK293
cells transfected with melanocortin MC1 receptor.
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3. Genotyped primary melanocytes in monoculture and co-culture
Fig. 4. Melanocyte co-culture p38 and p53 responses toNDP-MSHand ultraviolet radiation.
Co-cultures ofmelanocortinMC1 receptorwild type (WT—QF1193) or homozygous R151C melanocytes (QF1108) and keratinocytes were treated with 20 mJ/cm2 ultraviolet radiation= U, 20 nMNDP-MSH =N or both= UN and incubated for 1 hr.
C = Control, N = 20 nM NDP-MSH or F = 10 μM forskolin, for four days,
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4. Conclusion
in vitro studies using transfected cells orgenetically defined melanocyte monocultures and co-cultures have allcontributed to our knowledge of melanocortin MC1 receptor functionin human pigmentation and skin cancer risk.
It is becoming
increasingly apparent that the function of melanocortin MC1 recep-
tor
is much more complex than just regulating the switch from the red/
yellow pheomelanin to the brown/black eumelanin that was
originally described. Future melanocortin MC1 receptor pharmoco-
genetic
studies may utilise new transgenic mouse models.