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  • Title: Formation and removal of 8-MOP-DNA photoadducts in keratinocytes: effects of calcium concentration and retinoids.
    Author: Tokura Y, Edelson RL, Gasparro FP.
    Journal: J Invest Dermatol; 1991 Jun; 96(6):942-9. PubMed ID: 2045683.
    Abstract:
    8-methoxypsoralen (8-MOP)-DNA photoadducts were quantified in freshly isolated human and murine keratinocytes and cultured keratinocyte cell lines after in vitro treatment with 8-MOP (1-200 ng/ml) and ultraviolet A (UVA; 0.2-24.0 J/cm2). Greater doses of 8-MOP and UVA led to proportionately greater numbers of photoadducts, with a dose reciprocity relationship between the amounts of 8-MOP and UVA. No significant difference in photoadduct formation was observed between basal and differentiated cells. However, the transformed keratinocyte cell lines showed fewer photoadducts than did normal keratinocytes, which appeared to be correlated with the finding that the adduct formation was inhibited in normal keratinocytes cultured with phorbol 12-myristate 13-acetate, because this agent leads to epidermal hyperproliferation. In viable keratinocytes that were treated with a sublethal dose of 8-MOP and UVA (15 ng/ml and 1 J/cm2, respectively), 54% of photoadducts formed were removed over a 20-h period. Adduct removal depended on the calcium concentration in the media; cells cultured in standard high calcium levels showed a higher removal rate than those cultured in low-calcium media. The addition of retinoids (etretinate, acitretin, and 13-cis retinoic acid) to the culture induced 55 to 80% of suppression of the adduct removal. The calcium ionophore A23187 partially restored the suppression of photoadduct removal induced by retinoids. The present studies suggest that calcium performs an important role in the photoadduct removal and raise the possibility that the synergism of systemic retinoids and psoralen plus UVA photochemotherapy relates to the former's inhibition of repair of 8-MOP photoadducts in DNA.
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