Epigenetic immunomodulatory effect of eugenol and astaxanthin on doxorubicin cytotoxicity in hormonal positive breast Cancer cells

Figures

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  Fig. 1. Chemical structure of doxorubicin (a), eugenol (b) and astaxanthin (c)

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  Fig. 2. Effect of EUG (a), AST (b), DOX plus EUG (c) and DOX plus AST (d) on survival of MCF7 cells. Normalized isobologram constructed for the combination of increasing concentrations of DOX with decreasing concentrations of EUG (e) and AST (f). The combination index produced from 1 mM EUG combined with 0.25 μM DOX (g) and 40 μM AST combined with 0.25 μM DOX (h). Data are expressed as mean ± SD of three separate experiments, each performed in triplicates. a indicates significant change from control at P ≤ 0.05 using one-way ANOVA followed by Dunette as a post ANOVA test and b indicates significant from DOX alone at P ≤ 0.05 using student t-test.

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  Fig. 3. Effect of DOX, EUG, AST and their combination on Histones (H3 and H4) acetylation % (a), HAT protein expression (b), and % of HAT protein intensity normalised to β-actin (c). Data are expressed as mean ± SD of three separate experiments, each performed in triplicates. a,b,c and d indicate significant from control, DOX, EUG and AST, respectively at P ≤ 0.05 using one-way ANOVA followed by Tukey-Kramer as a post ANOVA test.

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  Fig. 4. Effect of DOX, EUG, AST and their combination on FOXP3, IFNγ and TNFα mRNA expression (a), aromatase and eGFR protein expression (), and % of aromatase and eGFR protein intensity normalised to β- actin (c). Data are expressed as mean ± SD of three separate experiments, each performed in triplicates. a,b,c and d indicate significant from control, DOX, EUG and AST, respectively at P ≤ 0.05 using one-way ANOVA followed by Tukey-Kramer as a post ANOVA test.

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  Fig. 5. Effect of DOX, EUG, AST and their combination on % of rhodamine123 uptake (a) and % of hoechst3342 uptake (b). Data are expressed as mean ± SD of three separate experiments, each performed in triplicates. a,b, c and d indicate significant from control, DOX, EUG and AST, respectively at P ≤ 0.05 using one-way ANOVA followed by Tukey-Kramer as a post ANOVA test.

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  Fig. 6. Effect of DOX, EUG, AST and their combination on % of cells at the phases of cell cycle (a), CK7 protein expression (b), and % of CK7 protein intensity normalised to β-actin (c). Data are expressed as mean ± SD of three separate experiments, each performed in triplicates. a,b, c and d indicate significant from control, DOX, EUG and AST, respectively at P ≤ 0.05 using one-way ANOVA followed by Tukey-Kramer as a post ANOVA test.

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  Fig. 7. Effect of DOX, EUG, AST and their combination on % of caspase 3 (a), mRNA expression of BAX, BCl2 and caspase 8 (b), % of cells according to propidium iodide staining (c), LC3I and LC3II protein expression (d), and LC3B II/I ratio (e). Data are expressed as mean ± SD of three separate experiments, each performed in triplicates. a,b, c and d indicate significant from control, DOX, EUG and AST, respectively at P ≤ 0.05 using one-way ANOVA followed by Tukey-Kramer as a post ANOVA test.