Figure 5

ZJW suppresses P-gp mediated MDR by inhibiting activation of NF-κB pathway in vitro . (A) ChIP analysis between NF-κB protein and ABCB1 gene. As a control, mouse monoclonal IgG was used. The blank group was the PCR results with no cDNA, and the input group was the cDNA from cell lysates without RIP procedure. (B) ChIP analysis was carried out to detect the effect of ABCB1 gene in HCT116/L-OHP cells treated with ZJW (50 μg/mL, 48 h). As a control, mouse monoclonal IgG was used. Input group was the cDNA from cell lysates without RIP procedure, and anti-NF-κB group was the cDNA from cell lysates after treatment with NF-κB antibody. (C) Western blotting assay was carried out to detect the level of P-gp, p-Akt (Ser473), p-IκB and p65 between HCT116 cell and HCT116/L-OHP cell. (D) The activity of the ABCB1 promoter in HCT116/L-OHP cells treated with LY294002 (20 μM, 2 h), ZJW (50 μg/mL, 48 h), and a combination of ZJW (50 μg/mL, 48 h) and IGF-1 (100 ng/mL, 48 h), was analyzed by a dual-luciferase assay kit. The results are the firefly luciferase/renilla luciferase ratio from different groups. Data are means ± standard deviation of values from triplicate experiments. **P < 0.01 vs. control group.