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. 2016 Mar 23;7(18):26003–26015. doi: 10.18632/oncotarget.8303

Figure 6. Huaier extract synergized with tamoxifen in signal pathway induction in ER-positive breast cancer cells.

Figure 6

A. Combined treatment induced more autophagy than either monotherapy (mTOR, p-mTOR, P70S6, p-P70S6, p-S6, P62, Atg7, Beclin1, and Lc3b). The autophagy effect was stronger in T47D than in MCF-7 cells. B. Western blot analysis revealed that Huaier extract synergizes with tamoxifen to induce apoptosis in ER-positive breast cancer cells (Cleaved-capase-3, Cleaved-capase-9, Cleaved-PARP, Bcl-2, and BCL2-associated X protein (BAX) proteins) and G0/G1 arrest (p-GSK3β, Cyclin D1). B, C, D. AKT is a target of the combination treatment. B. Effects of Huaier extract or/and TAM on expression of AKT and p-AKT proteins. C. Western blot analysis demonstrated that the transfection of AKT pcDNA3.1 reversed the protein changes induced by combined treatment (Beclin1, Lc3b, Bcl-2, Bax, and Cyclin D1). Expression of β-actin was used as an internal control in all above analyses. D. There was no significant difference in migration or invasion between AKT pcDNA3.1 with combined treatment and pcDNA3.1 without treatment groups. Bars, 50 μm. The data represent average cell numbers from at least 10 viewing fields. All results shown are from one of three independent experiments performed (*p < 0.05; **p < 0.01; ***p < 0.001 vs. the control group).