Gankyrin Drives Malignant Transformation of Gastric Cancer and Alleviates Oxidative Stress via mTORC1 Activation

Figure 4

Gankyrin alleviates oxidative stress in gastric cancer cell by activating mTORC1. (a) Both MKN45 and MKN74 cells with or without overexpressed gankyrin were treated with DMSO or 10 nM rapamycin for 1 h. The protein levels of pS6 K1, S6 K1, pS6, S6, p4E-BP1, and 4E-BP1 were analyzed by immunoblotting. (b) Oxidative stress was determined by measuring reactive oxygen species (ROS) levels using dihydroethidium (DHE) staining (red) in living cells. Representative images are shown (cell number > 100). Scale bars = 10 μm. (c) Quantification of fluorescence intensity in the experiment in (b). Data represent the means ± SD from three independent experiments in triplicate. represents a significant difference. (d) Oxidative stress was determined by measuring the levels of ROS using 2,7-dichlorodihydrofluorescein diacetate (H2DCF-DA) staining (green) in living cells. Representative images are shown (cell number > 100). Scale bars = 10 μm. (e) Quantification of fluorescence intensity in the experiment in (d). Data represent the means ± SD from three independent experiments in triplicate. represents a significant difference.

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