Xihuang Pill Induces Apoptosis of Human Glioblastoma U-87 MG Cells via Targeting ROS-Mediated Akt/mTOR/FOXO1 Pathway

Figure 6

The Akt/mTOR/FOXO1 signaling pathway was associated with the antigliomas of XHP. (a) After being incubated with different doses of XHP, 2 µM PTX or control medium for 24h, U-87 MG cells were lysed and subjected to western blot analysis of Akt, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 antibodies. (b) Western blot analysis of AKT, p-Akt, mTOR, p-mTOR, FOXO1, and p-FOXO1 expression in U-87 MG cells after treatment with LY294002 (20 μM), Rapamycin (50 nM), or NAC (5 mM) with or without XHP. (c) Confocal images of U-87 MG cells treated with XHP stained with anti-FOXO1 antibody. (d) Confocal images of U-87 MG cells treated with XHP were stained with anti-p-FOXO1 antibody. (e) Quantitative RT-PCR analysis of U-87 MG cells subjected to FOXO1-specific knockdown, results were normalized with GAPDH level and expressed as folds of control (n = 3, mean ± SD). (f) U-87 MG cells subjected to FOXO1-specific knockdown were assayed by western blot. (g) U-87 MG cells treated with XHP in the absence or presence of LY294002 (20 μM), Rapamycin (50 nM), or FOXO1-specific siRNA (10 nM). The proportion of Annexin V positive cells was measured by flow cytometry. Data are mean ± SD of three different experiments.P < 0.05, P < 0.01 significantly different from the control.

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