Journal: International Journal of Molecular Sciences

Article Title: The Impact of Inadequate Exposure to Epidermal Growth Factor Receptor–Tyrosine Kinase Inhibitors on the Development of Resistance in Non-Small-Cell Lung Cancer Cells

doi: 10.3390/ijms25094844

Figure Lengend Snippet: IC50 values for gefitinib in parental and gefitinib-resistant (GR) cell lines.

Article Snippet: EGFR-mutant (p.E746_A750del) NSCLC cell lines, HCC827 and PC9, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK), respectively.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Impact of Inadequate Exposure to Epidermal Growth Factor Receptor–Tyrosine Kinase Inhibitors on the Development of Resistance in Non-Small-Cell Lung Cancer Cells

doi: 10.3390/ijms25094844

Figure Lengend Snippet: Aberrant activation of EGFR signaling in GR cells. ( a , b ) Western blot analysis of EGFR expression and activation levels in HCC827 ( a ) and PC9 ( b ) parental and GR cells, in the absence or presence of gefitinib at the indicated concentrations. An anti-α-tubulin antibody was used for normalization. ( c , d ) Western blot analysis of expression and activation levels of ERK and AKT in HCC827 ( c ) and PC9 ( d ) parental and GR cells, in the absence or presence of gefitinib at the indicated concentrations. An anti-GAPDH antibody was used for normalization. Densitometric value ratios were calculated for phospho-ERK/total ERK (pERK/ERK) and phospho-AKT/total AKT (pAKT/AKT).

Article Snippet: EGFR-mutant (p.E746_A750del) NSCLC cell lines, HCC827 and PC9, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK), respectively.

Techniques: Activation Assay, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: The Impact of Inadequate Exposure to Epidermal Growth Factor Receptor–Tyrosine Kinase Inhibitors on the Development of Resistance in Non-Small-Cell Lung Cancer Cells

doi: 10.3390/ijms25094844

Figure Lengend Snippet: Evaluation of TGF-β1 expression in GR cells and its role in resistance and cell migration. ( a ) Immunofluorescence analysis of TGF-β1 expression (signaled in red) in HCC827 and PC9 parental and GR cell lines. Nuclei were stained with DAPI (signaled in blue). In the images, 20× magnification was used. ( b ) Immunoassay analysis of TGF-β1 levels in conditioned media from parental and GR cell lines. TGF-β1 levels were normalized for the cell number determined at the harvesting time and folds were calculated versus their respective parental cells. The values reported are the means from two independent experiments, each performed in duplicate. * p < 0.05 for comparison between parental versus GR cells (two-tailed Student’s t -test). ( c ) Cell proliferation assay on HCC827 parental and GR cells treated for 72 h with gefitinib (100 nM) and LY2109761 (LY, 5 µM), alone and in combination. *** p < 0.0005 for comparison between cells treated with gefitinib alone versus those treated with the drug combination (two-tailed Student’s t -test). ( d ) Analysis of wound-healing assays for HCC827 parental and GR-Low cells untreated and treated with LY (5 µM). *** p < 0.0005 for comparison between the treated versus untreated cells (two-tailed Student’s t -test).

Article Snippet: EGFR-mutant (p.E746_A750del) NSCLC cell lines, HCC827 and PC9, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK), respectively.

Techniques: Expressing, Migration, Immunofluorescence, Staining, Comparison, Two Tailed Test, Proliferation Assay