Fig. (6).

Ex vivo reduction of protein expressions of Bcl-2/Bax, c-Met, and NF-κB p-65 as well as phosphorylation (Ser⁴⁷³) of Akt in human breast cancer cells by serum from radiotherapy patient J taking EGCG for 8-w. Radiotherapy breast cancer patient J received oral dose of EGCG for 8-w, whereas radiotherapy patient C in control group did not receive EGCG. The blood samples were collected at 0-h at 0 w (immediately before EGCG administration), and at 2-h after EGCG administration on the 8th w. After MDA-MB-231 cells were treated with the 0 h sera (C0h, J0h) at 0 w, and 2-h sera (C8w, J8w) at 8-w from the patients, respectively, equal amounts of protein from total cell lysates were separated by SDS-PAGE, and Western blot analysis of Bcl-2/Bax (A), c-Met (B), NF-κB p-65 (C) and p-Akt (D) were done. For analysis of p-Akt, MDA-MB-231 cells were treated with each 0 h serum (C0h, J0h) at 0-w or 2-h serum at 8-w (C8w, J8w) from the patients, respectively. In the (A, B and C), the density of the band (normalized to β-actin) shown as mean ± SD is relative to that of the control C0h (designated as 1.00). In the (D), the density of the band (normalized to Akt) shown as mean ± SD is relative to that of the control C0h (designated as 1.00). For one experiment, 3 assays were carried out and only one set of gels is shown. In the (A, B and C), compared to the cells treated with the 0 h and 8-w sera from patient C in the control (C0h and C8w), the expression of Bax was significantly enhanced and the expressions of Bcl-2, c-Met and NF-κB were significantly reduced in MDA-MB-231 cells by the 8-w serum from the EGCG-treated patient J (J8w). In the (D), compared to the cells treated with the C0h and C8w sera from patient C in the control group, the phosphorylation (Ser⁴⁷³) of Akt in MDA-MB-231 cells was significantly reduced by the J8w serum from EGCG-treated patient J. Statistical analysis was carried out using the ANOVA and Bonferroni test. * P < 0.05 (n=3).