Article Figures & Data
Figures
- Figure 1.
WST-1 assay of three cell lines cultured with different concentrations of epidermal growth factor receptor tyrosine kinase inhibitors. At 48 h after exposure to gefitinib or erlotinib, the WST-1 assay was performed and the optical density was measured. Data are presented relative to the control (*p<0.01).
- Figure 2.
Extracellular-signal-regulated kinases (ERK) and AKT (protein kinase B) in A431 (A) and H23 (B) cells treated with epidermal growth factor receptor tyrosine kinase inhibitors for 6 h. After 24 h of pre-culture, EGFR-TKIs were added to the culture medium. Concentrations of gefitinib and erlotinib were 0.5 μM for A431 cells, and 20 μM for H23 cells (pERK: phospho-ERK, pAKT: phospho-AKT).
- Figure 3.
A: Cell-cycle analysis of H23 cells treated with epidermal growth factor receptor tyrosine kinase inhibitors. Cells were exposed to the indicated concentrations of gefitinib or erlotinib for 48 h. Collected cells were stained with propidium iodide and analyzed. B: Detection of apoptosis. H23 cells were exposed to staurosporine (0.2 μM, 2 h), gefitinib (20 μM, 48 h) or erlotinib (20 μM, 48 h). After incubation, collected cells were stained with 7-amino-actinomycin D (7-AAD) and annexin V-EnzoGold. A flow cytometer was used to analyze the fluorescence of samples. Low 7-AAD/low EnzoGold indicates living cells, high 7-AAD/low EnzoGold indicates necrotic cells, low 7-AAD/high EnzoGold indicates early apoptotic cells and high 7-AAD/high EnzoGold indicates late apoptotic cells.
- Figure 4.
A: Suppressive effects of gefitinib on caspase-3 activity in H23 cells. Cells were treated with 20 μM gefitinib for 48 h and harvested. Samples were reacted with substrates, and after incubation, absorbance at 405 nm was measured in order to determine the concentration of p-nitroaniline. Data show the ratio of activity in gefitinib-treated cells to those with DMSO. B: After treatment with 20 μM erlotinib, H23 cells were collected and the activity of caspase-3 was measured as describe above (*p=0.027, **p=0.018).
- Figure 5.
A: H23 cells were exposed to gefitinib or erlotinib for 48 h and harvested. For examination of expression of avian myelocytomatosis viral oncogene homolog (c-MYC), immunoblotting was performed. B: Reverse transcription polymerase chain reaction for c-MYC and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). After 48 h of treatment with gefitinib or erlotinib, cells were harvested (*1: p=0.03, *2: p=0.07, *3: p=0.38).
- Figure 6.
A: Cell-cycle analysis of H23 cells subjected to longer-term gefitinib treatment. Cells were exposed to 20 μM gefitinib for 72 h. Collected cells were stained with propidium iodide and analyzed. B: WST-1 assay of H23 cells treated with gefitinib for 3, 5 and 7 days. Data are relative to that of the control (*1 p<0.01, *2 p=0.02, *3 p<0.01).
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