Ginsenoside Rh4 Inhibits Colorectal Cancer Cell Proliferation by Inducing Ferroptosis via Autophagy Activation

Figure 2

Rh4 activates autophagy in human CRC cell lines. (a), (b) The viability of HT29 and HCT116 cells. HT29 and HCT116 cells were pretreated with 2 mM NAC (ROS scavenger), 2 mM 3-MA (autophagy inhibitor), 2 μM Fer-1 (ferroptosis inhibitor), 60 μM Z-VAD (apoptosis inhibitor), 40 μM Nec-1 (necrosis inhibitor), or 20 μM VX-765 (pyroptosis inhibitor) for 4 hours and then incubated with 200 μM Rh4 for 48 hours. (c) Effects of different concentrations of Rh4 or pretreatment with autophagy or ferroptosis before incubation with Rh4-containing medium on cell growth and morphology. Scale bars = 200 μm. (d), (e) and (f) Cells were treated with Rh4-containing medium or pretreated with autophagy inhibitors, and then Rh4-containing medium was added and incubated for 48 hours. The expression levels of autophagy-related proteins were determined by Western blotting. Values are presented as the means ± SD; n = 3; , , , compared with the control group (0 μM Rh4); ^ns#, ^##, ^### compared with the 200 μM Rh4 group.

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