Fig. 3

CHAG blocks activation of EGFR and its downstream signaling molecules, and inhibits EGF-induced expression of MMPs and PCNA in OC cells. a and b CHAG inhibited phosphorylation/activation of EGFR, Akt and ERK in A2780 and SKOV3 cells. The cells were cultured in serum free DMEM overnight and treated with 500 or 1000 μg/mL CHAG solutions for 1 h and then treated with 100 ng/ml EGF for 5 min. The cellular lysates were subjected to Western blotting. c and d Detection of the expression of MMP7 in A2780 and SKOV3 cells by Western blotting. In EGF group, the cells were treated with 100 ng/ml EGF for 24 h. In the CHAG + EGF groups, the cells were treated with 500 or 1000 μg/mL CHAG and 100 ng/mL EGF for 24 h. The cells were harvested and the lysates were subjected to Western blotting with anti-MMP7 antibodies. e and f Western blotting detection of the expression of PCNA in A2780 and SKOV3 cells. The cells were treated same as described in panel c and d. g and h CHAG blocked the activation of Rac1. The pull-down results were representatives of three independent experiments