Fig. 2

Effects of THC on apoptosis of breast cancer cells. A THC changed the mitochondrial membrane potential (ΔΨm) of breast cancer cells. MCF-7 and 4T1 cells were treated with various concentrations (0–80 μM) of THC for 48 h and then detect the change in ΔΨm by FCM. Data are expressed as means ± SD from three experiments (*P < 0.05; **P < 0.01; ***P < 0.001 compared to control). B MCF-7 and 4T1 cells were treated with designated concentrations (0–80 μM) of THC for 48 h. The harvested cells were loaded with DCFH-DA and then ROS levels in the cells were measured by FCM. Data are expressed as means ± SD from three experiments (*P < 0.05; **P < 0.01; ***P < 0.001compared to control). C Cell nuclear changes of MCF-7 and 4T1 cells were determined by staining with Hoechst 33258 and visualized by a fluorescence microscope after treatment with increasing doses of THC (0–80 μM) for 48 h. D MCF-7 and 4T1 cells were treated with THC (0–80 μM) at indicated doses for 48 h, and the level of apoptosis was evaluated using the Annexin V/PI dual-labeling technique,and determined by FCM. Data are expressed as means ± SD from three experiments (*P < 0.05; **P < 0.01; ***P < 0.001 compared to control). E The effect of THC (0, 80 μM) on Cleaved PARP protein expression was analyzed by immunofluorescence. F: Western blot analyses of MCF-7 and 4T1 cells treated (48 h) with different concentrations (0–80 μM) of THC were used to evaluate protein expression of Bax and Bcl-c, caspase-3, Cleaved capase-3, PARP and Cleaved PARP. Protein expression was quantified by the densitometry analysis using Image J and normalized against β-actin expression. Data are expressed as means ± SD from three experiments (*P < 0.05; **P < 0.01; ***P < 0.001 compared to control)