Research Article

Moscatilin Inhibits Lung Cancer Cell Motility and Invasion via Suppression of Endogenous Reactive Oxygen Species

Figure 5

Effect of moscatilin on endogenous reactive oxygen species (ROS) generation. H23 cells were treated with various nontoxic doses of moscatilin (0-1 μM) for various times (0–3 h). (a) Endogenous cellular ROS levels were determined by dichlorofluorescein diacetate (DCFH2-DA) probe. Values are mean ± SD ( ). versus nontreated control cells of each time point. (b) After the indicated treatment for 3 h, cells were incubated with hydroxyphenyl fluorescein (HPF) probe. Hydroxyl radical level was detected using fluorescence microplate reader. versus nontreated control cells. (c) Hydrogen peroxide level was examined using amplex red probe. versus nontreated control cells. (d) Superoxide anion level was detected by dihydroethidium (DHE) probe. versus nontreated control cells. (e) Cells were pretreated with 50 μM of ferrous sulfate (FeSO4) for 30 min prior to moscatilin treatments (0-1 μM) for 3 h. Endogenous ROS level were determined by using dichlorofluorescein diacetate (DCFH2-DA) probe. Values are mean ± SD ( ). versus nontreated control cells. versus ferrous sulfate treated cells. (f) Confluent monolayer of H23 cells was wounded using a 1 mm width tip and treated with moscatilin (1 μM) in the presence or absence of 50 μM of ferrous sulfate (FeSO4) for 24 h. Wound space was analyzed and represented as migration level relatively to the change of those in nontreated cells. versus nontreated control cells. versus ferrous sulfate treated cells.
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