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Abstract
Breast cancer is the most common malignancy in women, both in the developed and developing countries. Anthocyanins are natural coloring of a multitude of foods, such as berries, grapes or cherries. Glycosides of the aglycons delphinidin represent the most abundant anthocyanins in fruits. Delphinidin has recently been reported to inhibit the growth of human tumor cell line. Also, delphinidin is a powerful antioxidant that reportedly exerts beneficial effects in patients with advanced cancer by reducing the level of reactive oxygen species and increasing glutathion peroxidase activity. This study investigates the effects of delphinidin on protein ErbB2, ErbB3 and Akt expressions associated with cell proliferation and Bcl-2, Bax protein associated with cell apoptosis in MDA-MB-231 human breast cancer cell line. MDA-MB-231 cells were cultured with various concentrations (0, 5, 10, and 20 μ mol/L) of delphinidin. Delphinidin inhibited breast cancer cell growth in a dose dependent manner (p ＜ 0.05). ErbB2 and ErbB3 expressions were markdly lower 5 μ mol/L delphinidin (p ＜ 0.05). In addition, total Akt and phosphorylated Akt levels were decreased dose-dependently in cells treated with delphinidin (p ＜ 0.05). Futher, Bcl-2 levels were dose-dependently decreased and Bax expression was significantly increased in cells treated with delphinidin (p ＜ 0.05). In conclusion, I have shown that delphinidin inhibits cell growth, proliferation and induces apoptosis in MDA-MB-231 human breast cancer cell lines. (J Nutr Health 2013; 46(6): 503 – 510)
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Fig. 1.
Effect of delphinidin on cell proliferation in MDA-MB-231 cells. MDA-MB-231 cells were plated at a density of 2.5 × 104 cells/ml in 24 well plate with DMEM/F12 supplemented with 10% FBS for 48 hour, the monolayers were serum-starved with DMEM/F12 supplemented with 5 μ g/mL transferrin, 5 μ g/mL selenium, and 1 mg/mL bovine serum albumin for 24 hour. After serum starvation, the mo-nolayer were incubated in serum free medium with 0, 5, 10, 20 μ M delphinidin. Viable cell numbers were estimated by the MTT assay. Each bar represents the mean ± SE from three independent experiments. Comparison among different concentrations of delphinidin that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
Fig. 2.
Effects of delphinidin on ErbB2 expression in MDA-MB-231 cells. Cell lysates were subjected to immunoblotting with an antibody against ErbB2 and β-actin. A: The photographs of chemiluminescent detection of the blots, which were representative of three independent experiments are shown. B: Quantitative analysis of immunoblots. The relative abundance of each band was estimated by densitometric scanning of the exposed films. Each bar represents the mean ± SE (n = 3). Comparisons between groups that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
Fig. 3.
Effects of delphinidin on ErbB3 expression in MDA-MB-231 cells. MDA-MB-231 cells were treated with delphinidin for 48 hr as described in Fig. 1. Cell lysates were subjected to immunoblotting with an antibody against ErbB3 and β-actin. A: The photographs of chemiluminescent detection of the blots, which were representative of three independent experiments are shown. B: Quanti-tative analysis of immunoblots. The relative abundance of each band was estimated by densitometric scanning of the exposed films. Each bar represents the mean ± SE (n = 3). Comparisons between groups that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
Fig. 4.
Effects of delphinidin on Akt expression in MDA-MB-231 cells. Cells were treated as described in Fig. 1 and protein samples were analyzed by immunoblotting with anti-Akt protein antiboby or β-actin. A: The photographs of chemiluminescent detection of the blots, which are representative of three independent experiments, are shown. B: Quantitative analysis of immunoblots. The relative change in Akt band on Western blots was quantitated by densitometric analysis. Each bar represents the mean ± SE (n = 3). Comparisons between groups that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
Fig. 5.
Effects of delphinidin on p-Akt expression in MDA-MB-231 cells. Cells were treated as described in Fig. 1 and protein samples were analyzed by immunoblotting with antiphospho-Akt protein or β-actin antiboby. A: The photographs of chemiluminescent detection of the blots, which were representative of three independent experiments, are shown. B: Quantitative analysis of im-munoblots. The relative change in p-Akt band on western blots was quantitated by densitometric analysis. Each bar represents the mean ± SE (n = 3). Comparisons between groups that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
Fig. 6.
Effects of delphinidin on Bcl-2 expression in MDA-MB-231 cells. Cells were treated as described in Fig. 1 and protein samples were analyzed by immunoblotting with an Bcl-2 or β-actin antiboby. A: photographs of chemiluminescent detection of the blots, which were representative of threeindependent experiments, are shown. B: quantitative analysis of immunoblots. The relative abundance of each bar was estimated by densitometric scaning of the exposed films. Each bar represents the mean ± SE (n = 3). Comparisons between groups that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
Fig. 7.
Effects of delphinidin on Bax expression in MDA-MB-231 cells. Cells were treated as described in Fig. 1 and protein samples were analyzed by immunoblotting with an Bax or β-actin antiboby. A: photographs of chemiluminescent detection of the blots, which were representative of three independent experiments, are shown. B: quantitative analysis of immunoblots. The relative abundance of each bar was estimated by densitometric scaning of the exposed films. Each bar represents the mean ± SE (n = 3). Comparisons between groups that yielded significant differences (p ＜ 0.05) are indicated by different letters above each bar.
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