Comet assay demonstrates a higher ultraviolet B sensitivity to DNA damage in dysplastic nevus cells than in common melanocytic nevus cells and foreskin melanocytes.
We used the single cell gel electrophoresis assay (comet assay) to study ultraviolet B (UVB)-induced DNA damage in pigment cells. This assay detects DNA damage, mainly DNA strand breaks and alkali labile sites in the DNA molecule. We studied the effect of biologically relevant doses (comparable to 2-3 MED (minimal erythemal dose) for in vivo irradiated full-thickness skin) of monochromatic UVB light of 302 nm on cultured melanocytes derived from foreskin, common melanocytic nevi, and dysplastic nevi. We were able to demonstrate a linear dose-response relationship between UV dose and the migration coefficient of the comet tail in all three types of pigment cells. Nevus cells originating from dysplastic nevi showed the highest sensitivity to UVB irradiation: 65% higher induction of DNA damage compared to the induction in foreskin melanocytes. Common melanocytic nevus cells were most resistant and showed a 30% lower induction of DNA damage in comparison to foreskin melanocytes. Differences in chromatin structure and cell cycle profile may influence the results of the comet assay. Control experiments with x-ray irradiation, which is well known to produce direct DNA strand breaks via radical formation, revealed only small differences between the three types of melanocytic cells. It is unlikely, therefore, that intrinsic nuclear characteristics may account for the observed differences.
Noz
K C
KC
Department of Dermatology, University Hospital Leiden, The Netherlands.
Bauwens
M
M
van Buul
P P
PP
Vrolijk
H
H
Schothorst
A A
AA
Pavel
S
S
Tanke
H J
HJ
Vermeer
B J
BJ
D002465:Cell Movement; D004247:DNA / Q000528:radiation effects; D004249:DNA Damage; D004307:Dose-Response Relationship, Radiation; D004416:Dysplastic Nevus Syndrome / Q000235:genetics* / Q000473:pathology; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D008297:Male; D008544:Melanocytes / Q000502:physiology / Q000528:radiation effects*; D009508:Nevus, Pigmented / Q000235:genetics* / Q000473:pathology; D010413:Penis; D012878:Skin Neoplasms / Q000235:genetics* / Q000473:pathology; D014466:Ultraviolet Rays
D004247:DNA