Investigation of Acquired Resistance to EGFR-Targeted Therapies in Lung Cancer Using cDNA Microarrays

  • Kian Kani
  • Rafaella Sordella
  • Parag MallickEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 795)


Clinical tools to accurately describe, evaluate, and predict an individual’s response to cancer therapy are a field-wide priority; in many advanced cancers, only 10–20% of individuals will have a clinical benefit from therapy, yet we treat the entire population. Furthermore, many therapies are initially effective, but lose effectiveness over time. Here we describe methods to derive in vitro models of resistance to EGFR tyrosine kinase inhibitors. We additionally describe approaches to characterize possible mechanisms of resistance by genomic and transcriptomic approaches.

Key words

Gefitinib Erlotinib Non-small-cell lung cancer Therapeutic response Proteomics Resistance 


  1. 1.
    Scaltriti, M., and Baselga, J. (2006) The epidermal growth factor receptor pathway: a model for targeted therapy. Clin. Cancer Res. 12, 5268–5272.PubMedCrossRefGoogle Scholar
  2. 2.
    Lynch, T. J., Bell, D. W., Sordella, R., Gurubhagavatula, S., Okimoto, R. A., Brannigan, B. W., Harris, P. L., Haserlat, S. M., Supko, J. G., Haluska, F. G., Louis, D. N., Christiani, D. C., Settleman, J., and Haber, D. A. (2004) Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib. N. Engl. J. Med. 350, 2129–2139.PubMedCrossRefGoogle Scholar
  3. 3.
    Choi, S. H., Mendrola, J. M., and Lemmon, M. A. (2007) EGF-independent activation of cell-surface EGF receptors harboring mutations found in gefitinib-sensitive lung cancer. Oncogene 26, 1567–1576.PubMedCrossRefGoogle Scholar
  4. 4.
    Kumar, A., Petri, E. T., Halmos, B., and Boggon, T. J. (2008) Structure and clinical relevance of the epidermal growth factor receptor in human cancer. J. Clin. Oncol. 26, 1742–1751.PubMedCrossRefGoogle Scholar
  5. 5.
    Shigematsu, H., and Gazdar, A. F. (2006) Somatic mutations of epidermal growth factor receptor signaling pathway in lung cancers. Int. J. Cancer 118, 257–262.PubMedCrossRefGoogle Scholar
  6. 6.
    Engelman, J. A., Zejnullahu, K., Mitsudomi, T., Song, Y., Hyland, C., Park, J. O., Lindeman, N., Gale, C. M., Zhao, X., Christensen, J., Kosaka, T., Holmes, A. J., Rogers, A. M., Cappuzzo, F., Mok, T., Lee, C., Johnson, B. E., Cantley, L. C., and Janne, P. A. (2007) MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. Science 316, 1039–1043.PubMedCrossRefGoogle Scholar
  7. 7.
    Jiang, N., Leach, L. J., Hu, X., Potokina, E., Jia, T., Druka, A., Waugh, R., Kearsey, M. J., and Luo, Z. W. (2008) Methods for evaluating gene expression from Affymetrix microarray datasets. BMC Bioinformatics 9, 284.PubMedCrossRefGoogle Scholar
  8. 8.
    Kwak, E. L., Sordella, R., Bell, D. W., Godin-Heymann, N., Okimoto, R. A., Brannigan, B. W., Harris, P. L., Driscoll, D. R., Fidias, P., Lynch, T. J., Rabindran, S. K., McGinnis, J. P., Wissner, A., Sharma, S. V., Isselbacher, K. J., Settleman, J., and Haber, D. A. (2005) Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to gefitinib. Proc. Natl. Acad. Sci. USA 102, 7665–7670.PubMedCrossRefGoogle Scholar
  9. 9.
    Pao, W., Miller, V. A., Politi, K. A., Riely, G. J., Somwar, R., Zakowski, M. F., Kris, M. G., and Varmus, H. (2005) Acquired resistance of lung adenocarcinomas to gefitinib or erlotinib is associated with a second mutation in the EGFR kinase domain. PLoS Med. 2, e73.PubMedCrossRefGoogle Scholar
  10. 10.
    Engelman, J. A., Mukohara, T., Zejnullahu, K., Lifshits, E., Borras, A. M., Gale, C. M., Naumov, G. N., Yeap, B. Y., Jarrell, E., Sun, J., Tracy, S., Zhao, X., Heymach, J. V., Johnson, B. E., Cantley, L. C., and Janne, P. A. (2006) Allelic dilution obscures detection of a biologically significant resistance mutation in EGFR-amplified lung cancer. J. Clin. Invest. 116, 2695–2706.PubMedCrossRefGoogle Scholar
  11. 11.
    Oleykowski, C. A., Bronson Mullins, C. R., Godwin, A. K., and Yeung, A. T. (1998) Mutation detection using a novel plant endonuclease. Nucleic Acids Res. 26, 4597–4602.PubMedCrossRefGoogle Scholar
  12. 12.
    Qiu, P., Shandilya, H., D’Alessio, J. M., O’Connor, K., Durocher, J., and Gerard, G. F. (2004) Mutation detection using Surveyor nuclease. Biotechniques 36, 702–707.PubMedGoogle Scholar
  13. 13.
    Janne, P. A., Borras, A. M., Kuang, Y., Rogers, A. M., Joshi, V. A., Liyanage, H., Lindeman, N., Lee, J. C., Halmos, B., Maher, E. A., Distel, R. J., Meyerson, M., and Johnson, B. E. (2006) A rapid and sensitive enzymatic method for epidermal growth factor receptor mutation screening. Clin. Cancer Res. 12, 751–758.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.University of Southern CaliforniaLos AngelesUSA
  2. 2.Cold Spring Harbor LaboratoryCold Spring HarborUSA
  3. 3.Stanford UniversityStanfordUSA

Personalised recommendations