Fig. 4.

The effects of AKT activation on the inhibition of MDA-MB-231 and 4T1 cell proliferation and cell cycle arrest by Ezetimibe. (A) MDA-MB-231 and 4T1 cells were treated with different concentrations of SC79 (0, 0.5, 1, 5, 10, 15, 20, 40 μmol/L) for 48 h, and cell viability was measured using the CCK-8 assay. (B) MDA-MB-231 and 4T1 cells were treated with SC79 (0, 10 μmol/L) for 48 h, and the levels of t-AKT and p-AKT473 were detected using Western blotting analysis. (C) MDA-MB-231 cells were treated with different concentrations of Ezetimibe (0, 20, 40 μmol/L) and SC79 (0, 10 μmol/L) for 10 days, and 4T1 cells were treated for 14 days. The cell colony formation assay was used to measure the number of cell colonies formed. (D) MDA-MB-231 and 4T1 cells were treated with different concentrations of Ezetimibe (0, 20, 40 μmol/L) and SC79 (0, 10 μmol/L) for 48 h, and cell cycle distribution was analyzed using flow cytometry. (E) MDA-MB-231 and 4T1 cells were treated with different concentrations of Ezetimibe (0, 20, 40 μmol/L) and SC79 (0, 10 μmol/L) for 48 h, and the expression levels of Ki67, CDK2, CDK4, and CyclinD1 proteins were determined by Western blotting analysis. (F) MDA-MB-231 and 4T1 cells were treated with different concentrations of Ezetimibe (0, 20, 40 μmol/L) and SC79 (0, 10 μmol/L) for 48 h, and the levels of PDGFRβ, t-AKT, and p-AKT473 proteins were measured by Western blotting analysis. The results shown in the figure are representative of three independent experiments and are presented as mean ± standard deviation (Mean ± SD): *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Here 0 μmol/L Ezetimibe, SC79 or other chemicals refers to 0.1 % (v/v) DMSO solution.