Advertisement

Journal of Cancer Research and Clinical Oncology

, Volume 138, Issue 12, pp 2069–2077 | Cite as

Combined Erlotinib and Cetuximab overcome the acquired resistance to epidermal growth factor receptors tyrosine kinase inhibitor in non-small-cell lung cancer

  • Meng Wang
  • Jing Zhao
  • Lian-Min Zhang
  • Hui Li
  • Jin-Pu Yu
  • Xiu-Bao Ren
  • Chang-Li WangEmail author
Original Paper

Abstract

Purpose

Non-small-cell lung cancer (NSCLC) cells with somatic mutations in epidermal growth factor receptors (EGFR) are initially susceptible to tyrosine kinase inhibitor (TKI); however, eventually resistance to TKI is developed in these cells, which leads to the failure of treatment. The most common mechanism of this acquired drug resistance is development of a secondary T790M mutation in EGFR. In this study, we investigated the effects of the combination of Erlotinib and Cetuximab on T790M and L858R mutation lung cancer cells lines (H1975), in the primary NSCLC cells with the T790M mutation and TKI-resistant EGFR mutations human tumor xenograft model (H1975).

Methods

The effects of these two agents on cell proliferation, apoptosis, and EGFR-dependent signaling were evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, annexin V staining, and Western blotting. Sensitivity of EGFR inhibitors was detected in the primary tumor cell suspension and human tumor xenograft model (H1975).

Results

Compared with single-agent treatment, the combination of Cetuximab and Erlotinib increased apoptosis of EGFR TKI–resistant NSCLC cells (H1975), resulting in more pronounced growth inhibition on cell proliferation and significant inhibition of EGFR-dependent signaling.

Conclusions

These data suggest that treatment with a combination of Erlotinib and Cetuximab overcomes T790M-mediated drug resistance.

Keywords

NSCLC (non-small-cell lung cancer) Erlotinib Cetuximab T790M mutation Drug resistance Epidermal growth factor receptors (EGFR) 

Notes

Acknowledgments

We would like to thank Dr. Hong Yang and Rex Hadley for helping in editing this manuscript. We thank the Department of Immunology, Cancer Hospital of Tianjin Medical University for all technical assistance for the project. This study was supported by funding from the Science Foundations of Tianjin Medical University (2008ky21) and Tianjin Medical University Cancer Hospital (0906).

Conflict of interest

None.

References

  1. Barker AJ, Gibson KH, Grundy W et al (2001) Studies leading to the identification of ZD1839 (IRESSA): an orally active, selective epidermal growth factor receptor tyrosine kinase inhibitor targeted to the treatment of cancer. Bioorganic Med Chem Lett 11:1911–1914CrossRefGoogle Scholar
  2. Bean J, Riely GJ, Balak M et al (2008) Acquired resistance to epidermal growth factor receptor kinase inhibitors associated with a novel T854A mutation in a patient with EGFR-mutant lung adenocarcinoma. Clin Cancer Res 14:7519–7525PubMedCrossRefGoogle Scholar
  3. Brand TM, Dunn EF, Iida M, Myers RA, Kostopoulos KT, Li C, Peet CR, Wheeler DL (2011) Erlotinib is a viable treatment for tumors with acquired resistance to cetuximab. Cancer Biol Ther 12:436–446PubMedCrossRefGoogle Scholar
  4. Carey KD, Garton AJ, Romero MS et al (2006) Kinetic analysis of epidermal growth factor receptor somatic mutant proteins shows increased sensitivity to the epidermal growth factor receptor tyrosine kinase inhibitor, Erlotinib. Cancer Res 66:8163–8171PubMedCrossRefGoogle Scholar
  5. Chaft JE, Oxnard GR, Sima CS, Kris MG, Miller VA, Riely GJ (2011) Disease flare after tyrosine kinase inhibitor discontinuation in patients with EGFR-mutant lung cancer and acquired resistance to erlotinib or gefitinib: implications for clinical trial design. Clin Cancer Res 17:6298–6303PubMedCrossRefGoogle Scholar
  6. Fan Z, Lu Y, Wu X, Mendelsohn J (1994) Antibody-induced epidermal growth factor receptor dimerization mediates inhibition of autocrine proliferation of A431 squamous carcinoma cells. J Biol Chem 269:27595–27602PubMedGoogle Scholar
  7. Fukuoka M, Yano S, Giaccone G et al (2003) Multi-institutional randomized phase II trial of Gefitinib for previously treated patients with advanced non-small-cell lung cancer. J Clin Oncol 21:2237–2246PubMedCrossRefGoogle Scholar
  8. Han SW, Kim TY, Hwang PG et al (2005) Predictive and prognostic impact of epidermal growth factor receptor mutation in non-small-cell lung cancer patients treated with Gefitinib. J Clin Oncol 23:2493–2501PubMedCrossRefGoogle Scholar
  9. Jackman DM, Yeap BY, Sequist LV et al (2006) Exon 19 deletion mutations of epidermal growth factor receptor are associated with prolonged survival in non-small cell lung cancer patients treated with Gefitinib or Erlotinib. Clin Cancer Res 12:3908–3914PubMedCrossRefGoogle Scholar
  10. Jackman D, Pao W, Riely GJ, Engelman JA, Kris MG, Jänne PA, Lynch T, Johnson BE, Miller VA (2010) Clinical definition of acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors in non-small-cell lung cancer. J Clin Oncol 28:357–360PubMedCrossRefGoogle Scholar
  11. Janjigian YY, Groen HJ, Horn L, Smit EF, Fu Y, Wang F, Shahidi M, Denis LJ, Pao W, Miller VA (2011a) Activity and tolerability of afatinib (BIBW 2992) and cetuximab in NSCLC patients with acquired resistance to erlotinib or gefitinib. J Clin Oncol 29:7525Google Scholar
  12. Janjigian YY, Azzoli CG, Krug LM et al (2011b) Phase I/II trial of cetuximab and erlotinib in patients with lung adenocarcinoma and acquired resistance to erlotinib. Clin Cancer Res 17:2521–2527PubMedCrossRefGoogle Scholar
  13. Karashima T, Sweeney P, Slaton JW et al (2002) Inhibition of angiogenesis by the antiepidermal growth factor receptor antibody ImClone C225 in androgen-independent prostate cancer growing orthotopically in nude mice. Clin Cancer Res 8:1253–1264PubMedGoogle Scholar
  14. Kim HP, Han SW, Kim SH et al (2008) Combined lapatinib and Cetuximab enhance cytotoxicity against Gefitinib-resistant lung cancer cells. Mol Cancer Ther 7:607–615PubMedCrossRefGoogle Scholar
  15. Kobayashi H, Tanisaka K, Doi O et al (1997) An in vitro chemosensitivity test for solid human tumors using collagen gel droplet embedded cultures. Int J Oncol 11:449–455PubMedGoogle Scholar
  16. Kobayashi S, Boggon TJ, Dayaram T et al (2005a) EGFR mutation and resistance of non-small-cell lung cancer to Gefitinib. N Engl J Med 352:786–792PubMedCrossRefGoogle Scholar
  17. Kobayashi S, Ji H, Yuza Y et al (2005b) An alternative inhibitor overcomes resistance caused by a mutation of the epidermal growth factor receptor. Cancer Res 65:7096–7101PubMedCrossRefGoogle Scholar
  18. Kosaka T, Yatabe Y, Endoh H et al (2006) Analysis of epidermal growth factor receptor gene mutation in patients with non-small cell lung cancer and acquired resistance to Gefitinib. Clin Cancer Res 12:5764–5769PubMedCrossRefGoogle Scholar
  19. Kris MG, Natale RB, Herbst RS et al (2003) Efficacy of Gefitinib, an inhibitor of the epidermal growth factor receptor tyrosine kinase, in symptomatic patients with non-small cell lung cancer: a randomized trial. JAMA 290:2149–2158PubMedCrossRefGoogle Scholar
  20. Kurai J, Chikumi H, Hashimoto K et al (2007) Antibody-dependent cellular cytotoxicity mediated by Cetuximab against lung cancer cell lines. Clin Cancer Res 13:1552–1561PubMedCrossRefGoogle Scholar
  21. Kwak EL, Sordella R, Bell DW et al (2005a) Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to Gefitinib. Proc Natl Acad Sci USA 102:7665–7670PubMedCrossRefGoogle Scholar
  22. Kwak EL, Sordella R, Bell DW et al (2005b) Irreversible inhibitors of the EGF receptor may circumvent acquired resistance to Gefitinib. Proc Natl Acad Sci USA 102:7665–7670PubMedCrossRefGoogle Scholar
  23. Lynch TJ, Bell DW, Sordella R et al (2004) Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to Gefitinib. N Engl J Med 350:2129–2139PubMedCrossRefGoogle Scholar
  24. Matar P, Rojo F, Cassia R, Moreno-Bueno G, Di Cosimo S, Tabernero J, Guzmán M, Rodriguez S, Arribas J, Palacios J, Baselga J (2004) Combined epidermal growth factor receptor targeting with the tyrosine kinase inhibitor gefitinib (ZD1839) and the monoclonal antibody cetuximab (IMC-C225): superiority over single-agent receptor targeting. Clin Cancer Res 10:6487–6501PubMedCrossRefGoogle Scholar
  25. Mendelsohn J, Baselga J (2000) The EGF receptor family as targets for cancer therapy. Oncogene 19:6550–6565PubMedCrossRefGoogle Scholar
  26. Mitsudomi T, Kosaka T, Endoh H et al (2005) Mutations of the epidermal growth factor receptor gene predict prolonged survival after Gefitinib treatment in patients with non-small-cell lung cancer with postoperative recurrence. J Clin Oncol 23:2513–2520PubMedCrossRefGoogle Scholar
  27. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63Google Scholar
  28. Paez JG, Jänne PA, Lee JC et al (2004) EGFR mutations in lung cancer: correlation with clinical response to Gefitinib therapy. Science 304:1497–1500PubMedCrossRefGoogle Scholar
  29. Pao W, Chmielecki J (2010) Rational, biologically based treatment of EGFR mutant non-small-cell lung cancer. Nat Rev Cancer 10:760–774PubMedCrossRefGoogle Scholar
  30. Pao W, Miller VA (2005) Epidermal growth factor receptor mutations, small-molecule kinase inhibitors, and non-small-cell lung cancer: current knowledge and future directions. J Clin Oncol 23:2556–2568PubMedCrossRefGoogle Scholar
  31. Pao W, Miller V, Zakowski M et al (2004) EGF receptor gene mutations are common in lung cancers from “never smokers” and are associated with sensitivity of tumors to Gefitinib and Erlotinib. Proc Natl Acad Sci USA 101:13306–13311PubMedCrossRefGoogle Scholar
  32. Pao W, Miller VA, Politi KA et al (2005) Acquired resistance of lung adenocarcinomas to Gefitinib or Erlotinib is associated with a second mutation in the EGFR kinase domain. PLoS Med 2e:73CrossRefGoogle Scholar
  33. Pérez-Soler R, Chachoua A, Hammond LA et al (2004) Determinants of tumor response and survival with Erlotinib in patients with non-small-cell lung cancer. J Clin Oncol 22:3238–3247PubMedCrossRefGoogle Scholar
  34. Perez-Torres M, Guix M, Gonzalez A et al (2006) Epidermal growth factor receptor (EGFR) antibody down-regulates mutant receptors and inhibits tumors expressing EGFR mutations. J Biol Chem 281:40183–40192PubMedCrossRefGoogle Scholar
  35. Riely GJ, Pao W, Pham D et al (2006) Clinical course of patients with non-small cell lung cancer and epidermal growth factor receptor exon 19 and exon 21 mutations treated with Gefitinib or Erlotinib. Clin Cancer Res 12:839–844PubMedCrossRefGoogle Scholar
  36. Schaefer G, Shao L, Totpal K et al (2007) Erlotinib directly inhibits HER2 kinase activation and downstream signaling events in intact cells lacking epidermal growth factor receptor expression. Cancer Res 67:1228PubMedCrossRefGoogle Scholar
  37. Steiner P, Joynes C, Bassi R et al (2007) Tumor growth inhibition with Cetuximab and chemotherapy in non-small cell lung cancer xenografts expressing wild-type and mutated epidermal growth factor receptor. Clin Cancer Res 13:1540–1551PubMedCrossRefGoogle Scholar
  38. Traxler P, Bold G, Buchdunger E et al (2001) Tyrosine kinase inhibitors: from rational design to clinical trials. Med Res Rev 21:499–512PubMedCrossRefGoogle Scholar
  39. Wood ER, Truesdale AT, McDonald OB et al (2004) A unique structure for epidermal growth factor receptor bound to GW572016 (lapatinib): relationships among protein conformation, inhibitor off-rate, and receptor activity in tumor cells. Cancer Res 4:6652–6659CrossRefGoogle Scholar
  40. Yarden Y, Sliwkowski MX (2001) Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2:127–137PubMedCrossRefGoogle Scholar
  41. Ye D, Mendelsohn J, Fan Z (1999) Augmentation of a humanized anti-HER2 mAb 4D5 induced growth inhibition by a human-mouse chimeric anti-EGF receptor mAb C225. Oncogene 18:731–738PubMedCrossRefGoogle Scholar
  42. Zhang X, Gureasko J, Shen K et al (2006) An allosteric mechanism for activation of the kinase domain of epidermal growth factor receptor. Cell 125:1137–1149PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Meng Wang
    • 1
  • Jing Zhao
    • 2
  • Lian-Min Zhang
    • 1
  • Hui Li
    • 3
  • Jin-Pu Yu
    • 3
  • Xiu-Bao Ren
    • 3
  • Chang-Li Wang
    • 1
    Email author
  1. 1.Key Laboratory of Cancer Prevention and Therapy Tianjin, Department of Lung Cancer, Cancer Institute & HospitalTianjin Medical UniversityTianjinPeople’s Republic of China
  2. 2.Department of Breast Cancer, Cancer Institute & HospitalTianjin Medical UniversityTianjinPeople’s Republic of China
  3. 3.Department of Immunology, Cancer Institute & HospitalTianjin Medical UniversityTianjinPeople’s Republic of China

Personalised recommendations