Chlorogenic acid inhibits growth of 4T1 breast cancer cells through involvement in Bax/Bcl2 pathway

Objective(s): Chlorogenic acid is an herbal compound with various effects such as antiviral, antioxidant, and anticancer effect with low toxicity, which inhibits cell proliferation. Clinical studies had shown that chlorogenic acid has a positive effect on the different types of cancers treatment. Hence, this study evaluates chlorogenic acid effects on 4T1 breast cancer cells. Materials and Methods: In this study, cell proliferation was measured using an 3-(4,5-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assay (MTT) on 4T1 cells. Afterwards, other assays like P53, Caspase-3 proteins expression and Annexin V/PI were detected by flow cytometry. Also; Bax and Bcl-2 were carried out by immunocytochemistry. Results: 200 mu M of chlorogenic acid concentration showed the highest level of cytotoxicity toward 4T1 cells. Percentage of cell viability data were significant in 100 mu M (P < 0.05) and 150, 200 mu M (P < 0.001) doses. The evaluation using Annexin V/PI showed cell apoptosis in 100 mu M (P < 0.05), 150 mu M (P < 0.01), and for 200 mu M (P < 0.05 and P < 0.01). Immunocytochemistry results showed the upregulation of Bax and also the downregulation of Bcl-2 in 4T1 cells treated with chlorogenic acid (P < 0.001). The expression level of P53 and caspase-3 increased during treatment with chlorogenic acid in the 4T1 cells (P < 0.001). Conclusion: Our findings demonstrated that chlorogenic acid plays a notable role on apoptosis inducing in the 4T1 cells through regulation of apoptotic proteins.