Ultraviolet B injury increases prostaglandin synthesis through a tyrosine kinase-dependent pathway. Evidence for UVB-induced epidermal growth factor receptor activation

• Published in: The Journal of biological chemistry 1994-02, Vol.269 (5), p.3529-3533
• Main Authors: MILLER, C. C, HALE, P, PENTLAND, A. P
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Calcimycin - pharmacology; Catechols - pharmacology; Cell physiology; Cells, Cultured; Dinoprostone - biosynthesis; Effects of physical and chemical agents; Enzyme Activation; Epidermal Growth Factor - pharmacology; ErbB Receptors - metabolism; ErbB Receptors - radiation effects; Female; Fundamental and applied biological sciences. Psychology; Genistein; Humans; Isoflavones - pharmacology; Keratinocytes - enzymology; Keratinocytes - metabolism; Keratinocytes - radiation effects; Kinetics; Molecular and cellular biology; Nitriles - pharmacology; Protein-Tyrosine Kinases - antagonists & inhibitors; Protein-Tyrosine Kinases - metabolism; Signal Transduction - radiation effects; Skin - enzymology; Skin - metabolism; Skin - radiation effects; Tetradecanoylphorbol Acetate - pharmacology; Tyrphostins; Ultraviolet Rays
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(17)41895-3

To study the signal transduction mechanisms by which ultraviolet B (UVB) leads to increased prostaglandin E2 (PGE2) synthesis, human epidermal cultures were irradiated with 30 mJ/cm2 UVB and assayed for 6-h cumulative PGE2. Supernatants from irradiated cultures showed a 4-fold increase in PGE2 synthesis (113.6 +/- 26.8 pg/mg protein) when compared to supernatants from sham-irradiated cultures (25.6 +/- 3.9 pg/mg protein). Pretreatment of irradiated cultures with genistein (10 micrograms/ml) or tyrphostin-23 (50 microM), inhibitors of tyrosine kinases, blocked UVB-stimulated PGE2 synthesis. Treatment of nonirradiated cultures with epidermal growth factor (EGF), which acts through the receptor tyrosine kinase EGF-R, produced a 4-fold increase in PGE2 synthesis. However, addition of EGF to irradiated cultures did not further enhance their PGE2 synthesis, indicating irradiation rendered them refractory to EGF stimulation. In contrast, irradiated cultures could still significantly increase their PGE2 synthesis in response to the calcium ionophore A23187 or the protein kinase C activator 12-O-tetradecanoylphorbol-13-acetate, suggesting that the lack of response to EGF was selective. Furthermore, anti-phosphotyrosine immunoblot analysis revealed UVB-induced phosphorylation of tyrosine residues of EGF-R, an indicator of receptor activation. Phosphorylation was maximal 30-60 min after irradiation and was blocked by the tyrosine kinase inhibitors, genistein and tyrphostin. The antioxidant N-acetylcysteine decreased UVB-induced EGF-R tyrosine phosphorylation and PGE2 synthesis to near-basal levels. Conversely, treatment of unirradiated cultures with the potent oxidant tert-butyl-hydroperoxide (100 microM) increased both PGE2 synthesis and EGF-R phosphorylation. Collectively, these data suggest that antioxidant depletion induced by UV results in tyrosine phosphorylation and activation of the EGF-R. This activation may subsequently activate epidermal phospholipase at early time points after UVB exposure.