Research Paper Volume 12, Issue 7 pp 6240—6259

Inhibition of esophageal-carcinoma cell proliferation by genistein via suppression of JAK1/2-STAT3 and AKT/MDM2/p53 signaling pathways

Figure 5. Genistein inhibits protein phosphorylation and nuclear translocation. (A) The mRNA level of JAK1, JAK2, STAT1 and STAT3 were analyzed through qPCR in Eca-109 cells treated with different concentrations of genistein (2 μM, 4 μM, 8 μM) for 72 h. (B, C) The protein levels of JAK1, JAK2, STAT1, STAT3, p-JAK1, p-JAK2, p-STAT1 and p-STAT3 were measured through western blotting. (D) The mRNA levels of JAK1 and JAK2 in Eca-109 cells treated with 1 mM OV for 72 h. (E, F) The protein levels of JAK1, JAK2, p-JAK1 and p-JAK2 in Eca-109 cells treated with 1 mM OV for 72 h. (G) The mRNA levels of JAK1, JAK2 and STAT3 in Eca-109 cells treated with 1 mM OV or in combination with 4 μM genistein for 72 h. (H, I) The protein levels of JAK1, JAK2, STAT3, p-JAK1, p-JAK2 and p-STAT3 in Eca-109 cells treated with 1 mM OV or in combination with 4 μM genistein for 72 h. (J) The mRNA levels of Akt, MDM2 and Akt in Eca-109 cells treated with 8 μM genistein for 72 h. (K, L) The protein levels of P53, MDM2, p-MDM2, Akt and p-Akt in Eca-109 cells treated with 8 μM genistein for 72 h. (M, N) STAT3 and (O, P) MDM2 levels in the nucleus and cytoplasm of Eca-109 cells treated with 8 μM genistein for 72 h. All experiments were independently repeated three times. Data are analyzed using one-way ANOVA with Dunnett’s test and presented as the mean ± SD. *P<0.05; **P<0.01; ***P<0.001; Gen, genistein; OV, sodium orthovanadate; p-, phosphorylated.