| Purpose: Breast cancer is the leading cause of cancer-related death among women worldwide,and drug resistance remains a challenging in breast cancer treatment.In this study,we innovatively developed a new oleanolic acid derivative3-oxo-olean-12-en-28-acid(ZQL-1)and investigated the anti-tumor effect and related molecular mechanisms in breast cancer lines.Methods: Firstly,in vitro experiments were conducted using the Cell Counting Kit-8assay to test whether ZQL-1 inhibits the proliferation of MDA-MB-231,MCF-7,and SK-BR-3 cell lines,and to calculate the IC50 concentration of ZQL-1 in each cell line.Flow cytometry was used to detect apoptosis and cell cycle changes.The scratch wound healing assay was used to evaluate the effect of ZQL-1 on cell migration.Network pharmacology were used to predict ZQL-1 targets using the Pharm Mapper,Swiss Target Prediction,and Genecards database."cluster Profiler" R package were used to predict ralated pathway that may be affected by ZQL-1.Western blot analysis was used to detect changes in protein expression.Results: ZQL-1 showed concentration and time-dependent inhibition of proliferation in breast cancer cells MDA-MB-231,MCF-7,and SK-BR-3.The IC50 values of ZQL-1 in MDA-MB-231,MCF-7,and SK-BR-3 cells after 24 hours were 2.80μmol/L(95%CI:0.37-0.53),4.86μmol/L(95%CI: 0.578-0.79),and 3.7μmol/L(95%CI: 0.46-0.68),respectively.Compared with the control group,treatment with 0.5*IC50,IC50,and2*IC50 concentrations of ZQL-1 resulted in significant G0/G1 phase arrest in MCF-7cells(P<0.05),while the effect on the cell cycle in MDA-MB-231 and SK-BR-3 cells was not significant(P>0.05).With increasing concentrations of ZQL-1,the expression of cell cycle-related proteins CDK2,Cyclin D1,and Cyclin E1 was significantly reduced in MCF-7 cells(P < 0.05),but almost no effect was observed in MDA-MB-231 and SK-BR-3 cells(P>0.05).Compared with the control group,treatment with 0.5*IC50,IC50,and 2*IC50 concentrations of ZQL-1 resulted in a significant increase in the apoptosis rate of MDA-MB-231 and SK-BR-3 cells(P < 0.001),while no significant changes in apoptosis were observed in MCF-7 cells(P > 0.05).The ratio of apoptosis-related proteins cleaved caspase3/caspase 3,cleaved caspase7/caspase 7,and cleaved caspase9/caspase 9 showed a significant drug concentration-dependent increase in MDA-MB-231 and SK-BR-3 cells(P < 0.01),but no significant changes were observed in MCF-7 cells(P>0.05).Similarly,ZQL-1 inhibited the migration ability of MDA-MB-231 and SK-BR-3 cells(P<0.01),but had no significant effect on MCF-7cells(P>0.05).Network pharmacology analysis suggested the most likely mechanism of ZQL-1 is the EGFR mediated PI3K/AKT signaling pathway.Compared with the control group,treatment with 0.5*IC50,IC50,and 2*IC50 concentrations of ZQL-1 resulted in a concentration-dependent decrease in the p PI3K/PI3 K,p AKT/AKT,pm TOR/m TOR ratios,and e IF4 B protein expression in the three breast cancer cells(P < 0.05).In MDA-MB-231 and SK-BR-3 cells,the expression of 4EBP1 protein showed a concentration-dependent increase with ZQL-1 treatment(P < 0.05),while the p S6K1/S6K1 protein expression ratio significantly decreased with increasing ZQL-1concentration in MCF-7 cells(P<0.001).Conclusion: ZQL-1 inhibits the proliferation of breast cancer cells through the EGFR protein-mediated PI3K/AKT/m TOR pathway.In MDA-MB-231 and SK-BR-3 cells,ZQL-1 mainly exerts its anti-tumor proliferation effect by promoting apoptosis and inhibiting cell migration,while in MCF-7 cells,it mainly exerts its anti-tumor proliferation effect by blocking the cell cycle. |
PDF Full Text Request