Supplemental Fig. S1
The combination index of baicalein and docetaxel for cell viability in 8505c cells. The combination index was calculated from the Bliss independence model described by the equation CI=(EA+EB−[EA×EB])/EAB, where CI is the combination index, EA is the effect of drug A (baicalein), EB is the effect of drug B (docetaxel), and EAB is the combined effect of A and B at 24 hours (A) and 48 hours (B). aP<0.05; bP<0.01 vs. baicalein (20 µM) and docetaxel (2 nM).
enm-33-121-s001.pdf
Fig. 1Effects of the combined treatment with baicalein and docetaxel on the viability in 8505c cells. The cells were treated with baicalein (A, C) and docetaxel (B, D) individually or in combination (E, F) for 24 hours (A, B, E) and 48 hours (C, D, F). The cell viability was determined by MTT (thiazolyl blue tetrazolium bromide) assay. The data are presented as the mean±SEM of three independent experiments. aP<0.05; bP<0.1 vs. normal cells.
Fig. 2Effects of the combined treatment with baicalein and docetaxel on apoptosis in 8505c cells. The cells were treated with baicalein and docetaxel individually or in combination for 24 hours. (A) Nuclear fragmentation was determined by the observation of Hoechst staining using a fluorescence microscope (×400). The arrows indicate DNA fragmentation in apoptotic cells. Non-treated cells (a); baicalein-treated cells at 20 µM (b), 50 µM (c), and 100 µM (d); docetaxel-treated cells at 10 nM (e); cells treated the combination of docetaxel at 10 nM and baicalein at 20 µM (f), 50 µM (g), and 100 µM (h). The expression of Bax, caspase-3, cleaved caspase-3 (B), and Bcl-2 (C) proteins was measured by Western blotting analysis. The histogram (D, E, F, G) is presented as the mean±SEM of the band density for three independent experiments compared with β-actin expression. aP<0.05; bP<0.01 vs. baicalein alone and docetaxel combined treatment.
Fig. 3Effects of the combined treatment with baicalein and docetaxel on metastasis in 8505c cells. The cells were treated with baicalein and docetaxel individually or in combination for 24 hours. The expression of vascular endothelial growth factor (VEGF), transforming growth factor β1 (TGF-β1) (A), E-cadherin, and N-cadherin (D) proteins were measured by Western blotting analysis. The histogram (B, C, E, F) was presented as the mean±SEM of the band density for three independent experiments compared with β-actin expression. aP<0.01; bP<0.001 vs. baicalein alone and docetaxel combined treatment.
Fig. 4Effects of the combined treatment with baicalein and docetaxel on the signaling pathway in 8505c cells. The cells were treated with baicalein and docetaxel individually or in combination for 24 hours. The expression of mammalian target of rapamycin (mTOR) (A) and the phosphorylation of extracellular signal-regulated kinase (ERK) and Akt (B) were determined by Western blotting analysis. (C, D, E) Each histogram was presented as the mean±SEM of the band density for three independent experiments compared with β-actin expression (for mTOR) or the total expression (for ERK or Akt). aP<0.05; bP<0.01 vs. baicalein alone and docetaxel combined treatment.