Abstract: Objective To explore the new mechanism of Escin inhibiting the progression of breast cancer cells.Methods Escin treatment groups with different concentrations (0,10,20,30,40 μg/ml) were set up, and BC cells were treated with corresponding concentrations of Escin,then CCK8, clonal formation, flow cytometry, transmission electron microscopy and protein immunoblotting were used to evaluate the cell phenotype and possible mechanisms.Control group, Escin group and Escin+VX-765 group were set up,to determine the role of Caspase-1/GSDMD signaling pathway in Escin-induced pyroptosis of BC cells, cells were pretreated with Caspase-1 inhibitor VX-765.The cells in control group, Escin group and Escin+NAC group were pretreated with the reactive oxygen species (ROS) scavher N-Acetylcysteine (NAC),to determine the role of ROS in Escin induced pyroptosis of BC cells.Results Compared with the control group,different concentrations of Escin inhibited the proliferation and colony formation of BC cells in a concentration dependent manner (P<0.05).Compared with the control group,the ROS and pyroptosis rate were increased in Escin-treated group (P<0.05).The protein expression levels of FL-GSDMD and pro-Caspase-1 were significantly decreased in the Escin-treated group,while N-GSDMD,cleaved Caspase-1 and IL-18 protein expression were significantly increased (P<0.05).Compared with the Escin-treated group,the proliferation rate of Escin+VX-765 group was increased (P<0.05),and the expression of pyroptosis protein was decreased (P<0.05).Compared with the Escin-treated group,the proliferation rate of Escin+NAC group was increased (P<0.05),and the ROS,pyroptosis rate and pyroptosis protein expression were decreased (P<0.05).Conclusion The inhibitory effect of Escin on the progression of breast cancer cells may be related to its regulation of ROS/Caspase-1/GSDMD signaling pathway to promote cell pyroptosis.

Key words: breast cancer;pyroptosis;escin;GSDMD;caspase-1