ATR-Chk1 pathway inhibition promotes apoptosis after UV in primary human keratinocytes: potential basis for caffeine’s UV protective effects

摘要

Novel approaches to prevent and reverse ultraviolet (UV) damage are needed to combat rising sunlight-induced skin cancer rates. Mouse studies have shown that oral or topical caffeine promotes elimination of UV-damaged keratinocytes via apoptosis and markedly inhibits subsequent skin cancer development. This potentially important therapeutic effect has not been studied in human skin cells. Here we use primary human keratinocytes to examine which of several caffeine effects mediates this process. In these cells, caffeine more than doubled apoptosis after 75 mJ/cm² of UVB. Selectively targeting two of caffeine’s known effects did not alter UVB-induced apoptosis: inhibition of ATM (ataxia-telangiectasia mutated) and augmentation of cyclic AMP levels. In contrast, siRNA against ATR (ataxia-telangiectasia and Rad3-related) doubled apoptosis after UV through a p53-independent mechanism. Caffeine did not further augment apoptosis after UVB in cells in which ATR had been specifically depleted, suggesting a key target of caffeine in this effect is ATR. Inhibition of a central ATR target, Chk1, via siRNA or a novel highly specific inhibitor (PF610666) also augmented UVB-induced apoptosis. These data suggest that a relevant target of caffeine is the ATR-Chk1 pathway and that inhibiting ATR or Chk1 may have promise in preventing or reversing UV damage.