Hydroxyl safflower yellow B combined with doxorubicin inhibits the proliferation of human breast cancer MCF‑7 cells

Figure 1.. Combination of HSYB and DOX effectively inhibits the proliferation of breast cancer MCF-7 cells. Cellular viability was measured via the MTT method after 24 h of (A) DOX and (B) HSYB treatment at different concentrations. (C) Inhibition rate of DOX (0.4 µg/ml), HSYB (6 µg/ml) or DOX + HSYB combination treatment for 24 h. (D) Representative images of colony formation assays after DOX (0.4 µg/ml), HSYB (6 µg/ml) or DOX + HSYB treatment for 24 h. Data are presented as the mean ± SD (n=3). *P<0.05 and **P<0.01 vs. control; &&P<0.01 vs. DOX; ##P<0.01 vs. HSYB. DOX, doxorubicin; HSYB, hydroxyl safflower yellow B.

Figure 2.. Effects of combination of HSYB and DOX on apoptosis in breast cancer MCF-7 cells. (A) Morphological changes of cells were detected by Hoechst staining. Scale bar, 50 µm. (B) After Annexin V-FITC staining, flow cytometry was used to detect apoptosis in different groups. (C) Quantitative graph of MCF-7 cell apoptotic rate. Data are presented as the mean ± SD (n=3). **P<0.01 vs. control; &&P<0.01 vs. DOX; ##P<0.01 vs. HSYB. DOX, doxorubicin; HSYB, hydroxyl safflower yellow B; SSC, side scatter.

Figure 3.. Effects of combination of HSYB and DOX on the expression levels of apoptosis-associated molecular proteins in breast cancer MCF-7 cells. (A) BAX and BCL-2 protein expression were measured by western blotting. Quantitative graph of protein levels of (B) BCL-2, (C) BAX and (D) BCL-2/BAX in MCF-7 cells. (E) Protein expression levels of caspase-3, cleaved caspase-3, caspase-9 and cleaved caspase-9 were measured by western blotting. Quantitative graphs of protein expression levels of (F) cleaved caspase-3/total caspase-3 and (G) cleaved caspase-9/total caspase-9 in MCF-7 cells. Data are presented as the mean ± SD (n=3). **P<0.01 vs. control; &P<0.05 and &&P<0.01 vs. DOX; #P<0.05 and ##P<0.01 vs. HSYB. DOX, doxorubicin; HSYB, hydroxyl safflower yellow B.

Figure 4.. Effects of combination of HSYB and DOX on ROS levels in breast cancer MCF-7 cells. (A) ROS levels in MCF-7 cells were detected by flow cytometry. (B) Representative images of changes in intracellular ROS levels. Scale bar, 100 µm. (C) Changes in mitochondrial membrane potential in MCF-7 cells were detected by flow cytometry. (D) Quantitative graph of JC-1 red and green fluorescence ratios. (E) Cytoplasmic Cyto-c protein levels were measured by western blotting. (F) Quantitative graph of Cyto-c protein levels in MCF-7 cells. Data are presented as the mean ± SD (n=3). *P<0.05 and **P<0.01 vs. control; &P<0.05 vs. DOX; ##P<0.01 vs. HSYB. DOX, doxorubicin; HSYB, hydroxyl safflower yellow B; ROS, reactive oxygen species; Cyto-c, cytochrome c.