Modulation of lens epithelial cell proliferation by enhanced prostaglandin synthesis after UVB exposure

Abstract

Purpose: The goals of this investigation were to examine the synthesis of prostaglandins after UVB exposure of lens epithelial cells and to investigate their role in cell proliferation.

Methods: Cultured rabbit lens epithelial cells (cell line N/N1003A) were exposed to low levels of UV radiation. Prostaglandins were assayed by radioimmunoassay; products of arachidonic acid metabolism were analyzed by thin-layer chromatography and mass spectroscopy. Cell proliferation was measured by [3H]thymidine incorporation and proliferative autoradiography.

Results: Cultured lens epithelial cells exposed to UVB radiation showed a dose-dependent increase in basal prostaglandin synthesis measured 24 hours after UV exposure. At an optimal dose (250 J/m2) of UVB, prostaglandin E2 (PGE2) synthesis was enhanced tenfold. Product identity was confirmed using thin-layer chromatography and mass spectroscopy with authentic standards. Incubation of irradiated cells with exogenous arachidonic acid followed by extraction and thin-layer chromatography revealed that the cultures produced PGE2, prostaglandin I2 (measured as 6-keto-prostaglandin F1 alpha), prostaglandin F2 alpha, and hydroxyeicosatetraenoic acid. The synthesis of all of these products was enhanced threefold in cells exposed to 250-J/m2 UVB. Indomethacin pretreatment eliminated the synthesis of prostaglandins, further confirming their identity. To discover the relationship between PGE2 synthesis and irradiation-induced cell proliferation, [3H]thymidine incorporation into DNA was determined 24 or 48 hours after exposure. These experiments revealed a fivefold increase in incorporation induced by UVB exposure. UVB-enhanced incorporation of thymidine was eliminated by pretreatment of cultures with indomethacin to eliminate PG synthesis. However, when 100 nM PGE2 was added to the indomethacin-treated irradiated cultures, incorporation of the label was restored toward the level detected in the UVB-stimulated cells. Addition of other prostaglandins to the cultures was ineffective.

Conclusions: The results indicate that the synthesis of PGE2 is enhanced by exposure of lens epithelial cells to UVB radiation. PGE2 seems to play a specific role in cell proliferation after UV exposure. This increase in PGE2 synthesis may be important in posterior subcapsular cataract formation in humans and in animals exposed to UVB radiation in vivo.