Gigantol Inhibits Epithelial to Mesenchymal Process in Human Lung Cancer Cells

Figure 3

Effect of gigantol on epithelial to mesenchymal process (EMT) in human lung cancer cell H460. (a) H460 cells were treated with noncytotoxic doses of gigantol (0–20 μM) for 24 h. Wound space was photographed and analyzed at 0, 24, 48, and 72 h. The relative migration level was calculated as the changes of wound space of the treatment groups compared to that of the untreated control group at the indicated time. (b) H460 cells migration was examined using transwell migration assay. After 24 h the migrated cells were stained with Hoechst 33342 and visualized by fluorescence microscopy (scale bar is 50 μm). The relative migration level was calculated as the number of migrated cells of the treatment groups divided by that of the untreated control group. (c) H460 cells invasion was examined using transwell invasion assay. After 24 h the invaded cells were stained with Hoechst 33342 and visualized by fluorescence microscopy (scale bar is 50 μm). The relative invasion level was calculated as the number of migrated cells of the treatment groups divided by that of the untreated control group. (d) The effect of gigantol on migratory-related proteins. After H460 cells were treated with noncytotoxic doses of gigantol (0–20 μM) for 24 h, the expression of Rho GTP and Rac GTP was evaluated using Western blot assay. (e) The effect of gigantol on EMT marker proteins. After H460 cells were treated with noncytotoxic doses of gigantol (0–20 μM) for 24 h, the expressions of N-cadherin, E-cadherin, Vimentin, Snail, Slug, and ZEB-1 were evaluated using Western blot assay. The blots were reprobed with GAPDH to confirm equal loading. The immunoblot signals were qualified by densitometry. The data represent mean ± SE (). versus untreated control cells.

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