Effect of Lycorine on Trastuzumab Resistance in Breast Cancer Cells by Regulating AKT/mTOR Signaling Pathway
Lycorine has significant anti-tumor activity and can enhance the sensitivity and cytotoxicity of cisplatin to liver cancer cells.In this study,SKBR3 cells were cultured in vitro and induced to establish trastuzumab(TZB)resistant cells SKBR3/TR by continuous administration of 5 μg/mL TZB.The optimal concentration of lycorine was screened using CCK-8 method,and its effect on the TZB resistance index of SKBR3/TR cells was detected.A SKBR3/TR nude mouse model of transplanted tumors was constructed and randomly separated into a control group,TZB group,and TZB+lycorine group.After drug intervention,the tumor volume and mass of nude mice in each group were measured.SKBR3/TR cells cultured in vitro were randomly separated into control group,TZB group,TZB+SC79 group,TZB+lycorine group,and TZB+lycorine+SC79(AKT activator)group.After grouping and drug intervention,immunoblotting experiments were applied to detect the expression of AKT/mTOR signaling pathway related proteins.Flow cytometry,CCK-8 assay,and crystal violet staining were applied to detect apoptosis and proliferation of cells in each group.Immunoblotting experiments were applied to detect cell proliferation(PCNA),apoptosis(Bax,Cleaved Caspase-3,Bcl-2),and expression of resistance(P-glycoprotein(P-gp))related proteins in each group.The effect of lycorine on TZB resistance in breast cancer cells by regulating AKT/mTOR signaling pathway was investigated.The results show that lycorine could obviously reduce the TZB resistance index of SKBR3/TR cells and enhance the inhibitory effect of TZB on the growth of SKBR3/TR transplanted tumors in nude mice.Compared with the control group and TZB group,the apoptosis rate,expression of Bax and Cleaved Caspase-3 proteins in cells increased in the TZB+lycorine group(P＜0.05),the p-AKT/AKT,p-mTOR/mTOR,the cell viability,colony formation rate,expression of PCNA and Bcl-2,P-gp proteins decreased(P＜0.05);the trend of changes of various cellular indicators in the TZB+SC79 group was opposite to that in the TZB+Lycorianine group(P＜0.05).SC79 can weaken the effect of lycorine on SKBR3/TR cell lines.The results showed that lycorine can inhibit the activation of AKT/mTOR signaling pathway,thereby reducing the TZB resistance of breast cancer cells.
NETL
NSTL
万方数据
