Evidence-Based Complementary and Alternative Medicine / 2013 / Article / Fig 5

Research Article

Potential Therapeutic Role of Hispidulin in Gastric Cancer through Induction of Apoptosis via NAG-1 Signaling

Figure 5

Inhibition of NAG-1 expression and growth inhibition by ERK1/2 inhibitor. (a) AGS cells were treated with 30 μM Hispidulin in the presence or absence of the mitogen-activated protein kinase kinase 1/2 inhibitor PD98059, p38 inhibitor SB203580, and JNK1/2 inhibitor SP600125. For 48 h incubation, growth inhibition effect was determined by MTT assay. (b) AGS cells were treated with 30 μM Hispidulin for the indicated times. Total ERK1/2, phosphor-ERK1/2(pERK1/2), total p38, phosphor-p38(pp38), total JNK, and phosphor-JNK (pJNK) were detected by Western blot. (c) Attenuation of Hispidulin-induced NAG-1 upregulation in AGS cells by ERK inhibitor PD98059. AGS cells where treatment with 25–50 μM of PD98059 and NAG-1 expression was evaluated by Western blot analysis. (d) Apoptosis induced by Hispidulin was attenuated by PD98059 ERK inhibitor treatment followed by flow cytometry analysis. Each column represents the mean ± SD ( ; ; ). Expression of -actin was used as an internal control.