Abstract
A combination of 8-methoxypsoralen (8-MOP) and ultraviolet-A (UVA) radiation (320–400 nm) (PUVA) is widely used in the treatment of psoriasis and other skin diseases. PUVA is highly effective in eliminating hyperproliferative cells in the epidermis, but its mechanism of action has not been fully elucidated. In this study, we used immortalized JB6 mouse epidermal cells, p53−/−, and Fas ligand deficient (gld) mice to investigate the molecular mechanism by which PUVA induces cell death. The results indicate that PUVA treatment induces apoptosis in JB6 cells. In addition, PUVA treatment of JB6 cells results in p53 stabilization, phosphorylation, and nuclear localization as well as induction of p21Waf/Cip1 and caspase-3 activity. In vivo studies reveal that PUVA treatment induces significantly less apoptosis in the epidermis of p53−/− mice compared to p53+/+ mice. Furthermore, FasL-deficient (gld) mice are completely resistant to PUVA-induced apoptosis compared to wild-type mice. These results indicate that PUVA treatment induces apoptosis in mouse epidermal cells in vitro and in vivo and that p53 and Fas/Fas ligand interactions are required for this process, at least in vivo. This implies that similar mechanisms may be involved in the elimination of psoriatic keratinocytes from human skin following PUVA therapy.
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Abbreviations
- 8-MOP:
-
8-methoxypsoralen
- PUVA:
-
psoralen and UVA radiation
- CTCL:
-
cutaneous T-cell lymphoma
- FasL:
-
Fas Ligand
- MTT:
-
3-(4-5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- FACS:
-
fluorescence activated cell sorter
- TEM:
-
transmission electron microscopy
- TUNEL:
-
terminal deoxyribonucleotidyl transferase (TdT)-mediated dUTP nick end labeling
- FITC:
-
fluorescein isothiocyanate
- PI:
-
propidium iodide
- DMSO:
-
dimethylsulfoxide
- PEITC:
-
phenethyl isothiocyanate
- FADD:
-
Fas-associated death domain
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Acknowledgements
We thank S Bolshakov, K Ramirez, J Douglas, C Bucana, and K Dunner, Jr. for their excellent technical help, and M Goode and W Pagel for their excellent editorial assistance. This work was supported by National Cancer Institute grants CA 67174 (to HN Ananthaswamy) and CA 16672 (Institutional Core Grant). AB Santamaria and DW Davis were supported by predoctoral fellowships from the National Institutes of Health and National Institute of Environmental Health and Safety under training grants T32-CA-09598 and T32-ES-07290.
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Santamaria, A., Davis, D., Nghiem, D. et al. p53 and Fas ligand are required for psoralen and UVA-induced apoptosis in mouse epidermal cells. Cell Death Differ 9, 549–560 (2002). https://doi.org/10.1038/sj.cdd.4401007
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DOI: https://doi.org/10.1038/sj.cdd.4401007
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