Impacts of Leonurine on the malignant biological behaviors of breast cancer cells by regulating the Fas/FasL signaling pathway
Objective To investigate the impacts of Leonurine(Leo)on the proliferation,migration,apoptosis,cell cycle progression and epithelial mesenchymal transformation(EMT)of breast cancer cells and its mechanism.Methods 4T1 breast cancer cells were cultured in vitro.The cells were grouped into control group,low-dose Leonurine group(Leo-L),medium-dose Leonurine group(Leo-M),high-dose Leonurine group(Leo-H),high-dose Leonurine +transfection of short hairpin RNA lentivirus negative control group(Leo-H+sh NC group),and high-dose Leonurine +transfection of Fas short hairpin RNA lentivirus group(Leo-H+sh Fas group).The cells were treated with different concentrations of Leonurine(10-160mmol/L).CCK-8 assay was applied to detect the cell survival and to screen the optimal experimental concentration.Transwell assay was applied to evaluate the migration ability of cells.Flow cytometry was applied to detect cell apoptosis rate and cell cycle progression.Western blot was applied to detect the protein expressions of N-cadherin,Vimentin,fatty acid synthase(Fas)and fatty acid synthase ligand(FasL).A tumor-bearing mouse model was established to evaluate the effect of Leonurine on the growth of breast cancer xenografts.Results Treatment of Leonurine at the concentration of 10-160mmol/L for 12h significantly reduced the survival rate of 4T1 cells(P<0.05).The inhibitory concentration 50(IC50)value was(53.61±1.729)mmol/L,and the concentrations of 10,20,and 30mmol/L were selected for the subsequent experiments.Compared with those of the Control group,cells in the Leo-L,Leo-M and Leo-H groups presented significantly lower proliferative rate,migratory rate,cell ratio in the G2/M and S phase,and protein expressions of N-cadherin and Vimentin,but significantly higher apoptotic rate,cell ratio in the G0/G1 phase and protein expressions of Fas and FasL(P<0.05).Compared with those of Leo-H+sh-NC group,4T1 cells in the Leo-H+sh-Fas group presented significantly higher proliferative rate,migratory rate,cell ratio in the G2/M and S phase,and protein expressions of N-cadherin and Vimentin,but significantly lower apoptotic rate,cell ratio in the G0/G1 phase and protein expressions of Fas and FasL(P<0.05).Leonurine could inhibit the growth of breast cancer xenografts(P<0.05).Conclusion Leonurine can inhibit the malignant behaviors of breast cancer cells by activating the Fas/FasL signaling pathway.
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