Figure 7. The effects of genistein on EGFR expression and the reactivity of EsC cells to EGF. (A–C) EGFR expression in Eca-109 cells treated with different concentrations of genistein for 72 h was detected through qPCR and western blotting. EGFR expression in xenograft mice treated with (D–F) 5 mg/kg or (G–I) 10 mg/kg genistein for 42 d was significantly lower than that in an un-treated group. (J–L) EGFR expression in Eca-109 cells treated with 8 μM of genistein for 9 d. (M) The effect of recombined EGF (20 ng/mL) treatment alone on the proliferation of Eca-109 cells was detected through a CCK-8 assay. (N) The cell viability of Eca-109 cells incubated with genistein (8 μM) for 9 d following treatment with recombined EGF (20 ng/mL) for 24 h, 48 h or 72 h. (O) The reactivity of Eca-109 cells treated with genistein (8 μM) to recombined EGF. (P) A schematic working model to illustrate that genistein inhibits EGFR and its downstream pathways, the PI3K-Akt and JAK1/2-STAT3 signaling pathways, leading to apoptosis and cell cycle arrest in EsC cells. All in vitro experiments were independently repeated three times. The difference between two groups was tested using the Student’s t-test, and comparisons among multiple groups were performed using one-way ANOVA with Dunnett’s test. Data are presented as the mean ± SD. *P<0.05; **P<0.01; Gen, genistein.