Tuberc Respir Dis.  2014 Mar;76(3):120-126.

Apigenin and Wogonin Regulate Epidermal Growth Factor Receptor Signaling Pathway Involved in MUC5AC Mucin Gene Expression and Production from Cultured Airway Epithelial Cells

• Sikder MA
• Lee HJ
• Ryu J
• Park SH
• Kim JO
• Hong JH
• Seok JH
• Lee CJ

Abstract

BACKGROUND
We investigated whether wogonin and apigenin significantly affect the epidermal growth factor receptor (EGFR) signaling pathway involved in MUC5AC mucin gene expression, and production from cultured airway epithelial cells; this was based on our previous report that apigenin and wogonin suppressed MUC5AC mucin gene expression and production from human airway epithelial cells.
METHODS
Confluent NCI-H292 cells were pretreated with wogonin or apigenin for 15 minutes or 24 hours and then stimulated with epidermal growth factor (EGF) for 24 hours or the indicated periods.
RESULTS
We found that incubation of NCI-H292 cells with wogonin or apigenin inhibited the phosphorylation of EGFR. The downstream signals of EGFR such as phosphorylation of MEK1/2 and ERK1/2 were also inhibited by wogonin or apigenin.
CONCLUSION
The results suggest that wogonin and apigenin inhibits EGFR signaling pathway, which may explain how they inhibit MUC5AC mucin gene expression and production induced by EGF.

Keyword

Receptor, Epidermal Growth Factor; Mucins; Wogonin; Apigenin

MeSH Terms

Apigenin*
Epidermal Growth Factor*
Epithelial Cells*
Gene Expression*
Humans
Mucins*
Phosphorylation
Receptor, Epidermal Growth Factor*
Apigenin
Epidermal Growth Factor
Mucins
Receptor, Epidermal Growth Factor

Figure

• 

  Figure 1 Effect of wogonin or apigenin on proliferation of NCI-H292 cells. NCI-H292 cells were incubated for 72 hours in the presence of varying concentrations of wogonin or apigenin. Cell viability was determined using the sulforhodamine B assay as described in Materials and Methods. Each bar represents a mean±standard error of mean of three independent experiments in comparison with the control set at 100%. *Significantly different from control (p<0.05).

• 

  Figure 2 Effect of wogonin or apigenin on epidermal growth factor (EGF)-induced MUC5AC mucin production from NCI-H292 cells. NCI-H292 cells were pretreated with varying concentrations of wogonin or apigenin for 30 minutes and then stimulated with EGF (25 ng/mL) for 24 hours. Cell lysates were collected for measurement of MUC5AC mucin production by enzyme-linked immunosorbent assay. Three independent experiments were performed and the representative data were shown. Each bar represents a mean±standard error of mean of 3 culture wells in comparison with the control set at 100%. *Significantly different from control (p<0.05). †Significantly different from EGF alone (p<0.05).

• 

  Figure 3 Effect of wogonin or apigenin on epidermal growth factor receptor (EGFR) expression and phosphorylation. NCI-H292 cells were pretreated with wogonin (20 µM) or apigenin (20 µM) for 15 minutes and then stimulated with epidermal growth factor (EGF; 25 ng/mL) for 24 hours. Whole cell extract was collected and western blot analysis of the cellular proteins with anti-EGFR and anti-phospho-EGFR (Tyr1068) antibodies was conducted. Three independent experiments were performed and the representative data were shown.

• 

  Figure 4 Effect of wogonin or apigenin on phosphorylation of MEK1/2. NCI-H292 cells were pretreated with wogonin (20 µM) or apigenin (20 µM) for 24 hours and then stimulated with epidermal growth factor (EGF; 25 ng/mL) for the indicated periods. Whole cell extract was collected and western blot analysis of the cellular proteins with anti-phospho MEK1/2 (Ser221) antibody was conducted. Three independent experiments were performed and the representative data were shown.

• 

  Figure 5 Effect of wogonin or apigenin on phosphorylation of p44/42. NCI-H292 cells were pretreated with wogonin (20 µM) or apigenin (20 µM) for 24 hours and then stimulated with epidermal growth factor (EGF; 25 ng/mL) for the indicated periods. Whole cell extract was collected and western blot analysis of the cellular proteins with anti-phospho p44/42 (Thr202/Tyr204) antibody was conducted. Three independent experiments were performed and the representative data were shown.

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