Figure 2

The induction of necroptosis in osteosarcoma cells by shikonin. (A) K7, K12, K7M3, U2OS cells were treated with shikonin (3 μΜ) while 143B cells were treated with shikonin (6 μΜ) in the absence or presence of Nec-1 (50 μΜ) or Z-VAD-FMK (20 μΜ) for 8 hours. Nec-1 and Z-VAD-FMK were pretreated for 2 hours before shikonin. The cell survival rate was determined by MTT assay as described in Methods. ** represents p < 0.01. (B) K7 cells were treated with shikonin (3 μΜ) in the absence or presence of Nec-1 (50 μΜ) for 8 hours. Nec-1 was pretreated for 2 hours before shikonin. After treatment of shikonin, PI + cells increased significantly. When added with Nec-1 together with shikonin, the PI + cells induced by shikonin decreased significantly. Data are representative of 3 independent experiments.