Bufalin inhibits human breast cancer tumorigenesis by inducing cell death through the ROS-mediated RIP1/RIP3/PARP-1 pathways

Bufalin, a key active ingredient of the Chinese medicine Chan Su, inhibits breast cancer tumorigenesis in vitro and in vivo. Here, we found that the pan-caspase inhibitor zVAD-fmk failed to inhibit bufalin-induced cell death in MCF-7 and MDA-MB-231 human breast cancer cells, confirming that the cell death induced by bufalin is caspase-independent. Instead, bufalin increased the expression of the necroptosis mediators RIP1 and RIP3. Bufalin-induced cell death was prevented by small molecule inhibitors of RIP1 and poly (ADP-ribose) polymerase-1 (PARP-1) or genetic knockdown of RIP3 by shRNA transfection. In addition, ectopic RIP3 expression enhanced cell death by bufalin. We also found that bufalin increased intracellular reactive oxygen species levels; and cell death by bufalin was inhibited by the antioxidant NAC. In a mouse xenograft model of human breast cancer, bufalin induced PARP-1-dependent tumor cell death and inhibited tumor growth. These results demonstrated that bufalin inhibits human breast cancer tumorigenesis by inducing cell death through the reactive oxygen species-mediated RIP1/RIP3/PARP-1 pathways.