Effects of Baneh (Pistacia atlantica) Gum on Human Breast Cancer Cell Line (MCF-7) and Its Interaction with Anticancer Drug Doxorubicin

Abstract

Pistacia atlantica is one of the species of Anacardiaceae that grows in the wild in different regions of Iran. Traditionally, anacardiaceae family has antibacterial, fungicidal, and cytotoxic properties. Therefore, the present study was designed to investigate the possible cytotoxic and anti-proliferative properties of Baneh gum. Cytotoxicity of the plant gum was determined using MTT assay on MCF-7 human breast cancer cells. The cellular makers of apoptosis (caspase3 and P53) and cell proliferation (Cyclin-D1) were evaluated by western blotting. Doxorubicin was used as anticancer control drug in combination treatment. The result showed that Baneh gum (100 µg/mL) significantly induced cell damage, activated caspase3, and increased P53 protein level. In addition, Cyclin-D1 was significantly decreased in gum-incubated cells. Furthermore, combination treatment of cells with Baneh gum (25 µg/mL) and doxorubicin (200 nM) produced a significant cytotoxic effect as compared to each drug alone. In conclusion, Baneh gum (100 µg/mL) has a potential pro-apoptotic/anti-proliferative property against human breast cancer cells and its combination with doxorubicin in low doses may induce cell death effectively and be a potent modality to treat this type of cancer.

Keywords: Anti-apoptotic; Anti-proliferative; Baneh gum; MCF-7 and doxorubicin.

Figure 1

Effects of Baneh gum on MCF-7 cells viability. Cells were treated with gum for 24 h and then the viability was measured by MTT assay. Baneh gum reduced the cell viability, in dose dependent manner. Data are expressed as mean ±SEM; n = 6 wells for each group; **P < 0.01, and ***P < 0.001 significantly different versus control and vehicle treated cells

Figure 2

Effects of 25 µg/mL of Baneh gum alone or in combination with doxorubicin (200 nM) on MCF-7 cells. Combination treatment in comparison with each drug alone can induce significant cell toxicity. Each value in graph represents the mean ±SEM; n = 6 wells for each group; ***P < 0.001 significantly different versus control treated cells, +++P < 0.001 significantly different versus gum alone treated cells, #P < 0.05 significantly different versus doxorubicin alone incubated cells

Figure 3

Effect of Baneh gum treatment alone and in combination with 200 nM doxorubicin on caspase 3 protein activation. Cells were incubated with gum alone (25 µg/mL) or in combination with doxorubicin (200 nM) for 24 h and then proteins were extracted and protein expression was measured by western blot. Each value in graph represents the mean ± SEM band density ratio for each group; ***P < 0.001 significantly different versus control treated cells. β-actin was used as an internal control

Figure 4

Effect of Baneh gum treatment alone and in combination with 1µg/mLl doxorubicin on P53 protein level. Cells were incubated with gum alone (25 µg/mL) or in combination with doxorubicin (200 nM) for 24 h and then proteins were extracted and protein expression was measured by western blot. Each value in graph represents the mean ±SEM band density ratio for each group; **P < 0.01 significantly different versus control treated cells. β-actin was used as an internal control

Figure 5

Effect of Baneh gum treatment alone and in combination with 1µg/mL doxorubicin on Cyclin-D1 down-regulation. Cells were incubated with gum alone (25 µg/mL) or in combination with doxorubicin (200 nM) for 24 h and then proteins were extracted and protein expression was assayed by western blot. Each value in graph represents the mean ± SEM band density ratio for each group; **P< 0.01 significantly different versus control treated cells. β-actin was used as an internal control