The Synergistic Effects of Celastrol in combination with Tamoxifen on Apoptosis and Autophagy in MCF-7 Cells

Figure 5

Combination of celastrol and TAM triggers autophagy in MCF-7 cells. (a) Protein levels of LC3, P62, p-Akt, and p-mTOR were detected by western blot after treatment with celastrol (0.3 μM), TAM (1, 3, and 10 μM), or the combination for 24 hours in MCF-7 cells. The protein expression of LC3II increased significantly, the ratio of LC3II/U increased (), and the protein level of P62, an autophagy substrate marker, was also significantly reduced in the combination group. In addition, the protein expression of p-Akt and p-mTOR was repressed in the combination group. (b) MCF-7 cells were treated with celastrol (0.3 μM), tamoxifen (10 μM) alone, or the combination for 6 h. Cells were fixed and double-labelled with an anti-LC3 antibody (red), and nuclei were stained with DAPI (blue). An increase in red punctate aggregation in the cytoplasm indicated the activation of autophagy in the celastrol group and combination group. The white arrow points to an autophagosome.

(a)

(b)