The role of p38 in UVA-induced cyclooxygenase-2 expression in the human keratinocyte cell line, HaCaT.

Abstract

We examined the expression of cycloxygenase-2, the rate-limiting enzyme in the production of prostaglandins, in the UVA-irradiated human keratinocyte cell line, HaCaT. UVA induced a dose-dependent increase in COX-2 at the protein level at 2 and 4 h post-irradiation and at the mRNA level at 1 and 2 h post-irradiation. Experiments using semi-quantitative RT-PCR demonstrate that UVA increased the half-life of the COX-2 message by more than fourfold in the presence of Actinomycin D (with a half life between 4 and 8 h post-irradiation), suggesting that UVA induction of COX-2 is post-transcriptionally regulated. Through the use of the specific p38 inhibitor, SB202190, increases in COX-2 message and protein levels were abrogated in UVA-irradiated cells. In UVA-irradiated cells treated with SB202190, the half-life of the COX-2 message was decreased to basal levels (between 1 and 2 h post-irradiation), indicating that p38 was responsible for the stabilization of the message. Luciferase activity was increased in UVA-irradiated cells transfected with reporter constructs containing the 3' UTR of COX-2, a region containing AU-rich elements (AREs). These regulatory sequences of AUUUA have been proposed as one mechanism of post-transcriptional regulation. Increases observed in luciferase activity could be decreased using a p38 dominant-negative construct. We report for the first that UVA can induce COX-2 expression in the human keratinocyte cell line, HaCaT. Additionally, p38 appears to play a critical role in the UVA-induced expression of COX-2 in these keratinocytes and may serve as a potential drug target in the chemoprevention of skin cancer.