Anthricin Isolated from Anthriscus sylvestris (L.) Hoffm. Inhibits the Growth of Breast Cancer Cells by Inhibiting Akt/mTOR Signaling, and Its Apoptotic Effects Are Enhanced by Autophagy Inhibition
Figure 4
Anthricin inhibits Akt/mTOR signaling. (a) Anthricin inhibits the phosphorylation of mTOR, S6K1, and Akt in a dose-dependent manner. MCF-7 cells were treated with varying concentrations of the drug for 12 h or 100 nM rapamycin for 1 h, and the phosphorylation states of mTOR (p-Ser2448), S6K1 (p-Thr389), and Akt (p-Ser473) were analyzed by immunoblotting. (b) Anthricin inhibits the phosphorylation of mTOR, S6K1, and Akt in a time-dependent manner. MCF-7 cells were treated with 25 nM anthricin for the time period specified. (c) Anthricin is a potential target of mTOR and Akt. MCF-7 cells were transduced with TSC2 siRNA or scrambled siRNA (control). Cells were treated with or without 25 nM anthricin, and the proteins were analyzed by immunoblotting. (d) Anthricin disrupts the association between mTOR and raptor. mTOR immunoprecipitates were isolated from anthricin-treated MCF-7 cells by using an anti-mTOR antibody, and the amounts of raptor and mTOR in the immune complexes were analyzed by immunoblotting.
