Melatonin Inhibits Reactive Oxygen Species-Driven Proliferation, Epithelial-Mesenchymal Transition, and Vasculogenic Mimicry in Oral Cancer

Figure 3

Melatonin abated the migration and invasion of oral cancer cell by inhibiting ROS-activated Akt signaling. (a–d) Cal27 and FaDu cells were exposed to melatonin (1 mM), NAC (5 mM), or the vehicle (control) for 24 h. Transwell assays were performed to assess the (a) migration and (c) invasion of Cal27 and FaDu cells. The number of (b) migrated cells and (d) invaded cells was calculated. (e) Cal27 and FaDu cells were exposed to melatonin (1 mM), LY294002 (20 μM), NAC (5 mM), or the vehicle (control) for 24 h. Immunofluorescence staining was conducted to analyze the expression of E-cadherin, Vimentin, and Snail. Magnification: 100x. (f) Cal27 and FaDu cells were treated with melatonin (1 mM) or vehicle (control) for 24 h. Western blot was performed to detect the expressions of p-Akt, Akt, E-cadherin, Vimentin, and Snail. GAPDH was used as endogenous control. (g) Cal27 and FaDu cells were treated with LY294002 (20 μM), NAC (5 mM), or the vehicle (control) for 24 h. Representative Western blot results of p-Akt, Akt, Snail, E-cadherin, and Vimentin. GAPDH was used as endogenous control. The ratios from the indicated proteins to GAPDH are indicated below the bands. Data are represented as the mean ± SD of three independent experiments. versus control group. Ctrl: control; Mel: melatonin; NAC: N-acetyl-L-cysteine.

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