Figure 6

Effect of genistein (Gen) on TPA-induced EGFR, MAPK, IκB, AKT, PKC-signaling pathways in hepatoma cell lines. (A) Cells were pretreated with 10 or 20 μM Gen for 30 min before incubation with 80 μM TPA for 45 min or 24 h (for EGFR), then whole-cell lysates were prepared and subjected to western blotting with antibodies specific for phosphorylated PI3k, AKT, IκB, JNK, p38, and ERK as described in Methods section. (B) HepG2 cells were cultured in serum-free media containing 80 μM TPA for 45 min, then the cell cytosolic and membrane fractions were prepared and then the extracts were analyzed by western blots as described in Methods section. The values under each lane indicate relative density of the band normalized to β-actin. All analyses were representative of at least three independent experiments. (C) The gelatinolytic activities of MMP-9 were determined by gelatin zymography as described in Materials and Methods.