Physiological doses of ultraviolet irradiation induce DNA strand breaks in cultured human melanocytes, as detected by means of an immunochemical assay.
An immunochemical assay, i.e. sandwich enzyme-linked immunosorbent assay, has been modified to detect UV-induced damage in cellular DNA of monolayer-grown human melanocytes. The method is based on the binding of a monoclonal antibody to single-stranded DNA. The melanocytes derived from human foreskin of skin type II individuals were suspended and exposed to UVA, UVB, solar-simulated light or gamma-rays. Following physiological doses of UVA, UVB or solar-simulated light, a dose-related DNA unwinding comprising a considerable number of single-strand breaks (ssb) was observed. No correlation was found between different seeded cell densities or different culturing periods and the UVA sensitivity of the cells. After UVA irradiation, 0.07 ssb/10(10) Da/kJ/m2 were detected and after UVB irradiation 1.9 ssb/10(10) Da/kJ/m2 were seen. One minimal erythema dose of solar-simulated light induced 2.25 ssb/10(10) Da. Our results from melanocytes expressed in ssb/Da DNA are comparable and have the same sensitivity toward UVA as well as toward UVB as nonpigmented skin cells. As low doses of UVA have already been shown to induce detectable numbers of ssb, this assay is of great interest for further investigations about the photoprotecting and/or photosensitizing effects of melanins in human melanocytes derived from different skin types.
Wenczl
E
E
Department of Dermatology, Kun Street Hospital, Budapest, Hungary.
Pool
S
S
Timmerman
A J
AJ
van der Schans
G P
GP
Roza
L
L
Schothorst
A A
AA
D002478:Cells, Cultured; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007231:Infant, Newborn; D008544:Melanocytes / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects*; D014466:Ultraviolet Rays
D004247:DNA