Figure 2. TE inhibits cell proliferation, PGE2 synthesis, COX mRNA expression and migration.
(A) Line EpH4, 66.1 or MCF-7 cells were seeded at 2.5×105 cells/well/1.0 ml media in triplicate in 24 well plates. PBS or TE (6.25, 12.5, 25 µl) was added. Forty-eight hours later, cell number was determined. (* P <0.05).
(B) 6× 106 of line 410.4 cells were seeded in 12 ml media in triplicate. TE from a stock concentration of 2µg/µl was added to the media in the indicated amounts. Control dishes contained PBS at the highest amount used with TE. Twenty-four hours later, cell-conditioned media was collected and PGE2 level was determined by EIA. Data expressed as pg PGE2/ mg protein as mean±SE. (P <0.05). (C,D) 6× 106 line 410.4 cells were seeded in 12 ml media and 24 hours after plating, TE was added as in B. Six hours later RNA was isolated, reverse transcribed, and amplified using primer specific for COX-2 (C) and COX-1 (D). (E) Line 410.4 tumor cells, untreated or pretreated with TE (6.25–50 µg/ml) for 1 hour were placed in the upper chamber of a modified Boyden assay. Media or media containing 2% serum was added to the lower chamber. Cells traversing the membrane were quantified 24 hours later.