Effects of 2’nitroflavone on the EGF signaling pathway and cell proliferation in breast cancer cells

CEBRÓN JR; BOJORGE MA; MARDER M; MIQUET JG ; GONZÁLEZ L

Mar del Plata

Congreso; LXVII Reunión anual de la Sociedad Argentina de Investigaciones Clínicas (SAIC); 2022

Sociedad Argentina de Investigaciones Clínicas

Epidermal growth factor receptor (EGFR) is a tyrosine kinase receptor associated with tumorigenesis of several tissues and has been proposed to be involved in the molecular mechanism of action of flavonoids. For that reason, flavonoids are proposed to be combined with drugs that target this receptor.2’nitroflavone (2’NF) is a synthetic flavone obtained in our institute that has previously demonstrated to produce apoptosis in breast cancer cells and affect the expression of receptors related to EGFR activity. With the aim of investigating if a combinatory therapy involving 2’NF and EGFR inhibitors would be a possible effective treatment for breast cancer, the present study analyzes if this flavone modulates the EGF signaling pathway and cell proliferation activity. MDA-MB-231 and MCF-7 breast cancer cells were treated with 2NF at a concentration of 20 µM and 10 µM respectively or vehicle for 1 and 48 h. Afterward, cells were stimulated with EGF (25 ng/ml) for 10 min. The protein content and phosphorylation at activating residues of EGFR, Akt and Erk were assessed by immunoblotting. Besides, the effect on cell proliferation was determined by hexosaminidase assay. For this determination MDA-MB-231 and MCF-7 cells were seeded and treated with 2’NF (20 µM and 10 µM respectively) or vehicle and EGF (25 ng/ml) for 48 h. Results showed a reduction in EGFR phosphorylation induced by EGF after 48 h of treatment with 2’NF in both cell lines (p0.05); while in the case of Akt there is a diminution in its phosphorylation after 1 (p0.05) and 48 h (p0.01). In the case of Erk phosphorylation there is a decrease in MCF-7 (p0.01) but an increase in MDA-MB-231 (p0.05). As regards EGF-induced proliferation, a decrease was observed in both cell lines (p0.0001) when cells were treated with 2’NF. In conclusion, 2’NF demonstrated to have effects on EGF signaling pathway and cell proliferation activity which could justify a combinatory therapy involving 2’NF and EGFR inhibitors.