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J.J. Cools-Lartigue, E. Lewandowski, T.S. Chernin, J.A. Marshall, E. Antecka, M.N. Burnier Jr.; The Expression of Prostaglandin E2 (PGE2) and Interleukin 6 (IL-6) in Macrophage and Uveal Melanoma Conditioned Media . Invest. Ophthalmol. Vis. Sci. 2003;44(13):1560.
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Purpose: The relation between tumor associated macrophages (TAM) and tumor cells in uveal melanoma has not been elucidated. Cytokines, such as IL-6 and Prostaglandin E2, are involved in tumor/TAM interactions, and have been shown to be poor prognostic markers in uveal and cutaneous melanoma. This study investigated the levels of IL-6 and PGE2 in macrophage and uveal melanoma conditioned media. Methods: Five human uveal melanoma cell lines (92.1, MKT-BR, OCM-1, SP6.5), one transformed melanocytic cell line (UW-1), and one macrophage cell line (28SC) were seeded in RPMI-1640 medium at a concentration of 1x106 cells/ml. After 18 hours, RPMI diluted (20%) media from the uveal melanoma cell lines was placed on the macrophage cell line. Diluted macrophage media was placed on each uveal melanoma cell line. Tumor and macrophages without conditioned media were used as controls. IL-6 and PGE2 levels were determined by ELISA and EIA respectively prior to media transfer and 6, 12, 24, and 36 hours thereafter. Results: In the absence of conditioned media both PGE2 and IL-6 levels were negative. In the presence of conditioned media neither macrophages nor uveal melanoma cells expressed PGE2. IL-6 was upregulated in all cells following the addition of conditioned media. Expression of IL-6 appeared at 6 hours in macrophages, and at 36 hours in the uveal melanoma cells (19.01ug/ml). Macrophages incubated with tumor cell media showed a greater increase in IL-6 (50.38>300.00ug/ml) than tumor cells incubated with macrophage media (~0ug/ml). Conclusions: PGE2 expresssion remained negligible after exposure to conditioned media. IL-6 expression was increased in both uveal melanoma and macrophages after treatment with conditioned media. However, macrophages began expressing IL-6 earlier and at much higher levels than uveal melanoma cells. We postulate that the upregulation of IL-6 was not due to the expression of PGE2 and is mediated by an unknown secondary pathway.
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