DOI: 10.5061/dryad.nvx0k6f1f
title: Data from: Oncogenic transformation by inhibitor-sensitive and resistant EGFR mutants

description: Background   Somatic mutations in the kinase domain of the
 epidermal growth factor receptor tyrosine kinase gene EGFR are common in
 lung adenocarcinoma. The presence of mutations correlates with tumor
 sensitivity to the EGFR inhibitors erlotinib and gefitinib, but the
 transforming potential of specific mutations and their relationship to
 drug sensitivity have not been described.   Methods and Findings
   Here, we demonstrate that EGFR active site mutants are
 oncogenic. Mutant EGFR can transform both fibroblasts and lung epithelial
 cells in the absence of exogenous epidermal growth factor, as evidenced by
 anchorage-independent growth, focus formation, and tumor formation in
 immunocompromised mice. Transformation is associated with constitutive
 autophosphorylation of EGFR, Shc phosphorylation, and STAT pathway
 activation. Whereas transformation by most EGFR mutants confers on cells
 sensitivity to erlotinib and gefitinib, transformation by an exon 20
 insertion makes cells resistant to these inhibitors but more sensitive to
 the irreversible inhibitor CL-387,785.   Conclusion  
 Oncogenic transformation of cells by different EGFR mutants causes
 differential sensitivity to gefitinib and erlotinib. Treatment of lung
 cancers harboring EGFR exon 20 insertions may therefore require the
 development of alternative kinase inhibition strategies.
description: # Data from: Oncogenic transformation by inhibitor-sensitive and resistant
 EGFR mutants
 [https://doi.org/10.5061/dryad.nvx0k6f1f](https://doi.org/10.5061/dryad.nvx0k6f1f) ## Description of the data and file structure These representative photos are fields from a set of soft agar assay plates in which NIH-3T3 cells ectopically expressing wild-type or mutant EGFR are suspended in soft agar to quantify oncogenic potential and measure response to EGFR inhibitors.  Because wild-type EGFR expressing cells are not oncogenically transformed, EGF has been added to stimulate EGFR signaling and soft agar colony formation in order to measure inhibitor response.  This experiment is an independent validation of the conclusions in the PLoS Medicine paper (doi [10.1371/journal.pmed.0020313](https://doi.org/10.1371/journal.pmed.0020313) ), for which the raw image data no longer exist.  This experiment was performed within a few months of the published experiment in 2005. ### Files and variables #### File: 20050707\_Iressa\_\_Tarceva\_\_and\_HKI\_soft\_agar\_photos.zip **Description:**  This folder contains data from an experiment performed in 2005.  The first three subfolders, labeled "A1L858R"*, *"ins124 exon 20 insertion", and "WT+EGF" contain soft agar photo data collected from three NIH-3T3 cell lines, each expressing EGFR L858R, EGFR D770_N771insNPG, or wild-type EGFR (with exogenous EGF added).  Inside each of these cell type folders are another 3 folders, each labeled with the inhibitor used to treat the cells.  HKI = HKI-272, Iressa = gefitinib, and Tarceva = erlotinib.  Within each of these inhibitor folders are a set of 6 folders, each labeled with the concentration of inhibitor used to treat the cells, ranging from 0 to 10 uM.  We ran out of HKI-272 and could not complete the 10 uM dose set for EGFR L858R expressing cells and wild-type EGFR expressing cells, and these folders are labeled to reflect the lack of inhibitor treatment.  Within each dose defined folder are a set of (usually) 30 images acquired at random across the plate.  In samples for which no transformed cells survived and no colonies were observed (at the highest doses of inhibitor), only 3 photos (usually) were taken at random across the plate. ## Code/software None,Soft agar photos were quantified manually. # Funding MZ is supported by the National Institutes of Health-sponsored MD-PhD program. WCH is supported by the Doris Duke Charitable Foundation and the National Cancer Institute. WRS and MM are supported by the Novartis Research Foundation, the Claudia Adams Barr Foundation, and the Charles A. Dana Human Cancer Genetics Program of the Dana-Farber Cancer Institute, the Poduska Family Foundation, the Damon-Runyon Cancer Research Foundation, Joan's Legacy, the American Cancer Society, the Flight Attendant Medical Research Institute, and the National Cancer Institute.