Description(s):
Epidermal growth factor receptor (EGFR) is a cell-surface receptor with intrinsic intracellular protein-tyrosine kinase (TK) activity. Ligand binding induces EGFR dimerization and phosphorylation, leading to the activation of the EGFR signaling pathway. In several malignancies such as non-small cell lung cancer (NSCLC), EGFR signaling is deregulated due to mutations in EGFR, which results in uncontrolled proliferation and migration of tumor cells. EGFR mutations can lead to oncogene-addicted cancers, where the tumor cells depend on the mutated EGFR for cell survival and the malignant phenotype. One of the most common EGFR mutations found in human patients is L858R substitution in exon 21, within the activation loop of EGFR. Patients with this mutation are sensitive to EGFR tyrosine kinase inhibitors (TKIs) such as gefitinib or erlotinib, whereas patients with wild-type EGFR are not sensitive to TKI. Another clinically relevant mutation associated with acquired gefitinib and erlotinib resistance is T790M, found in exon 20. Cells expressing EGFR with both L858R and T790M mutations are resistant to induced apoptosis in the presence of gefitinib or erlotinib.
 
This Ba/F3 cell line was established by electroporating parental Ba/F3 cells with a vector containing the wt EGFR gene under an MSCV promoter along with GFP and hygromycin resistance. Following transfection, clones with resistance to hygromycin were subsequently screened for GFP expression, with protein expression of the mutant EGFR tested via western blot. The clone with the highest expression were selected and expanded to produce this stable cell line.