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 3
Autophagy plays a cytoprotective role in anthricin-treated breast cancer cells. (a and b) Autophagy inhibition promotes cell death. Cells were treated with 25 nM anthricin and/or 25 µM CQ for 12 h. Data are expressed as a percentage of the control and shown as mean ± SD (). compared to vehicle-treated cells. (c) Cotreatment of anthricin and CQ is more sensitive to the apoptotic cell death in MCF-7 and MDA-MB-231. Apoptosis was analyzed by flow cytometry and annexin V staining as described in Materials and Methods. Data are represented as means ± S.D. as determined from 3 independent experiments. compared to anthricin alone. (d) Anthricin enhances cell death in ULK1 or Atg13 knockdown cells. MCF-7 cells stably transduced with lentiviral shRNA were treated with anthricin (25 nM) for 12 h. Data are expressed as a percentage of the control and shown as the mean ± SD (). compared with shGFP control cells within the same concentration. (e) Apoptosis increased in ULK1- and Atg13-silenced MCF-7 cells after anthricin treatment. The extent of apoptosis in cells treated with 25 nM anthricin for 12 h was analyzed by immunoblotting. (f) Anthricin is more sensitive to the apoptotic cell death in ULK1-silenced MCF-7 cells. Apoptosis was analyzed by flow cytometry and annexin V staining. compared with shGFP control cells within the same concentration.
