Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

The role of cooperativity with Src in oncogenic transformation mediated by non-small cell lung cancer-associated EGF receptor mutants

Oncogene volume 28, pages 1821–1832 (2009)Cite this article

Abstract

Non-small cell lung cancer (NSCLC)-associated epidermal growth factor receptor (EGFR) mutants are constitutively active and induce ligand-independent transformation in non-malignant cell lines. We investigated the possibility that the ability of mutant EGFRs to transform cells reflects a constitutive cooperativity with Src using a system in which the overexpression of mutant, but not wild-type, EGFR induced anchorage-independent cell growth. Src was constitutively activated and showed enhanced interaction with mutant EGFRs, suggesting that constitutive EGFR–Src cooperativity may contribute to mutant EGFR-mediated oncogenesis. Indeed, the mutant EGFR-mediated cell transformation was inhibited by Src- as well as EGFR-directed inhibitors. Importantly, a tyrosine to phenylalanine mutation of the major Src phosphorylation site on EGFR, Y845, reduced the constitutive phosphorylation of NSCLC-EGFR mutants, as well as that of STAT3, Akt, Erk and Src, and reduced the mutant EGFR–Src association as well as proliferation, migration and anchorage-independent growth. Reduced anchorage-independent growth and migration were also observed when dominant-negative-Src was expressed in mutant EGFR-expressing cells. Overall, our findings show that mutant EGFR–Src interaction and cooperativity play critical roles in constitutive engagement of the downstream signaling pathways that allow NSCLC-associated EGFR mutants to mediate oncogenesis, and support the rationale to target Src-dependent signaling pathways in mutant EGFR-mediated malignancies.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8

Similar content being viewed by others

References

  • Alvarez JV, Greulich H, Sellers WR, Meyerson M, Frank DA . (2006). Signal transducer and activator of transcription 3 is required for the oncogenic effects of non-small-cell lung cancer-associated mutations of the epidermal growth factor receptor. Cancer Res 66: 3162–3168.

    Article  CAS  Google Scholar 

  • Biscardi JS, Maa MC, Tice DA, Cox ME, Leu TH, Parsons SJ . (1999). c-src-mediated phosphorylation of the epidermal growth factor receptor on Tyr845 and Tyr1101 is associated with modulation of receptor function. J Biol Chem 274: 8335–8343.

    Article  CAS  Google Scholar 

  • Brom ann PA, Korkaya H, Courtneidge SA . (2004). The interplay between src family kinases and receptor tyrosine kinases. Oncogene 23: 7957–7968.

    Article  CAS  Google Scholar 

  • Di Fiore PP, Pierce JH, Fleming TP, Hazan R, Ullrich A, King CR et al. (1987). Overexpression of the human EGF receptor confers an EGF-dependent transformed phenotype to NIH 3T3 cells. Cell 51: 1063–1070.

    Article  CAS  Google Scholar 

  • Dimri M, Naramura M, Duan L, Chen J, Ortega-Cava C, Chen G et al. (2007). Modeling breast cancer-associated c-src and EGFR overexpression in human MECs: C-src and EGFR cooperatively promote aberrant three-dimensional acinar structure and invasive behavior. Cancer Res 67: 4164–4172.

    Article  CAS  Google Scholar 

  • Fu YN, Yeh CL, Cheng HH, Yang CH, Tsai SF, Huang SF et al. (2008). EGFR mutants found in non-small cell lung cancer show different levels of sensitivity to suppression of src: implications in targeting therapy. Oncogene 27: 957–965.

    Article  CAS  Google Scholar 

  • Golas JM, Arndt K, Etienne C, Lucas J, Nardin D, Gibbons J et al. (2003). SKI-606, a 4-anilino-3-quinolinecarbonitrile dual inhibitor of src and abl kinases, is a potent antiproliferative agent against chronic myelogenous leukemia cells in culture and causes regression of K562 xenografts in nude mice. Cancer Res 63: 375–381.

    CAS  PubMed  Google Scholar 

  • Greulich H, Chen TH, Feng W, Janne PA, Alvarez JV, Zappaterra M et al. (2005). Oncogenic transformation by inhibitor-sensitive and -resistant EGFR mutants. PLoS Med 2: e313.

    Article  Google Scholar 

  • Gschwind A, Fischer OM, Ullrich A . (2004). The discovery of receptor tyrosine kinases: targets for cancer therapy. Nat Rev Cancer 4: 361–370.

    Article  CAS  Google Scholar 

  • Haura EB, Zheng Z, Song L, Cantor A, Bepler G . (2005). Activated epidermal growth factor receptor-stat-3 signaling promotes tumor survival in vivo in non-small cell lung cancer. Clin Cancer Res 11: 8288–8294.

    Article  CAS  Google Scholar 

  • Herbst RS . (2004). Review of epidermal growth factor receptor biology. Int J Radiat Oncol Biol Phys 59: 21–26.

    Article  CAS  Google Scholar 

  • Ishizawar R, Parsons SJ . (2004). c-src and cooperating partners in human cancer. Cancer Cell 6: 209–214.

    Article  CAS  Google Scholar 

  • Kazansky AV, Rosen JM . (2001). Signal transducers and activators of transcription 5B potentiates v-src-mediated transformation of NIH-3T3 cells. Cell Growth Differ 12: 1–7.

    CAS  PubMed  Google Scholar 

  • Kim H, Chan R, Dankort DL, Zuo D, Najoukas M, Park M et al. (2005). The c-src tyrosine kinase associates with the catalytic domain of ErbB-2: Implications for ErbB-2 mediated signaling and transformation. Oncogene 24: 7599–7607.

    Article  CAS  Google Scholar 

  • Kloth MT, Laughlin KK, Biscardi JS, Boerner JL, Parsons SJ, Silva CM . (2003). STAT5b, a mediator of synergism between c-src and the epidermal growth factor receptor. J Biol Chem 278: 1671–1679.

    Article  CAS  Google Scholar 

  • Lombardo LJ, Lee FY, Chen P, Norris D, Barrish JC, Behnia K et al. (2004). Discovery of N-(2-chloro-6-methyl- phenyl)-2-(6-(4-(2-hydroxyethyl)- piperazin-1-yl)-2-methylpyrimidin-4- ylamino)thiazole-5-carboxamide (BMS-354825), a dual Src/Abl kinase inhibitor with potent antitumor activity in preclinical assays. J Med Chem 47: 6658–6661.

    Article  CAS  Google Scholar 

  • Lynch TJ, Bell DW, Sordella R, Gurubhagavatula S, Okimoto RA, Brannigan BW 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: 2129–2139.

    Article  CAS  Google Scholar 

  • Maa MC, Leu TH, McCarley DJ, Schatzman RC, Parsons SJ . (1995). Potentiation of epidermal growth factor receptor-mediated oncogenesis by c-src: implications for the etiology of multiple human cancers. Proc Natl Acad Sci USA 92: 6981–6985.

    Article  CAS  Google Scholar 

  • Maeda M, Shintani Y, Wheelock MJ, Johnson KR . (2006). Src activation is not necessary for transforming growth factor (TGF)-beta-mediated epithelial to mesenchymal transitions (EMT) in mammary epithelial cells. PP1 directly inhibits TGF-beta receptors I and II. J Biol Chem 281: 59–68.

    Article  CAS  Google Scholar 

  • Masaki T, Igarashi K, Tokuda M, Yukimasa S, Han F, Jin YJ et al. (2003). Pp60c-src activation in lung adenocarcinoma. Eur J Cancer 39: 1447–1455.

    Article  CAS  Google Scholar 

  • Mitsudomi T, Kosaka T, Endoh H, Horio Y, Hida T, Mori S 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: 2513–2520.

    Article  CAS  Google Scholar 

  • Muthuswamy SK, Muller WJ . (1995). Direct and specific interaction of c-src with neu is involved in signaling by the epidermal growth factor receptor. Oncogene 11: 271–279.

    CAS  PubMed  Google Scholar 

  • Paez JG, Janne PA, Lee JC, Tracy S, Greulich H, Gabriel S et al. (2004). EGFR mutations in lung cancer: correlation with clinical response to gefitinib therapy. Science 304: 1497–1500.

    Article  CAS  Google Scholar 

  • Pao W, Miller VA, Politi KA, Riely GJ, Somwar R, Zakowski MF 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 2: e73.

    Article  Google Scholar 

  • Politi K, Zakowski MF, Fan PD, Schonfeld EA, Pao W, Varmus HE . (2006). Lung adenocarcinomas induced in mice by mutant EGF receptors found in human lung cancers respond to a tyrosine kinase inhibitor or to down-regulation of the receptors. Genes Dev 20: 1496–1510.

    Article  CAS  Google Scholar 

  • Reddi AL, Ying G, Duan L, Chen G, Dimri M, Douillard P et al. (2007). Binding of cbl to a phospholipase Cgamma1-docking site on platelet-derived growth factor receptor beta provides a dual mechanism of negative regulation. J Biol Chem 282: 29336–29347.

    Article  CAS  Google Scholar 

  • Sharma SV, Bell DW, Settleman J, Haber DA . (2007). Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer 7: 169–181.

    Article  CAS  Google Scholar 

  • Shimamura T, Lowell AM, Engelman JA, Shapiro GI . (2005). Epidermal growth factor receptors harboring kinase domain mutations associate with the heat shock protein 90 chaperone and are destabilized following exposure to geldanamycins. Cancer Res 65: 6401–6408.

    Article  CAS  Google Scholar 

  • Shtiegman K, Kochupurakkal BS, Zwang Y, Pines G, Starr A, Vexler A et al. (2007). Defective ubiquitinylation of EGFR mutants of lung cancer confers prolonged signaling. Oncogene 26: 6968–6978.

    Article  CAS  Google Scholar 

  • Silva CM . (2004). Role of STATs as downstream signal transducers in src family kinase-mediated tumorigenesis. Oncogene 23: 8017–8023.

    Article  CAS  Google Scholar 

  • Song L, Morris M, Bagui T, Lee FY, Jove R, Haura EB . (2006). Dasatinib (BMS-354825) selectively induces apoptosis in lung cancer cells dependent on epidermal growth factor receptor signaling for survival. Cancer Res 66: 5542–5548.

    Article  CAS  Google Scholar 

  • Sordella R, Bell DW, Haber DA, Settleman J . (2004). Gefitinib-sensitizing EGFR mutations in lung cancer activate anti-apoptotic pathways. Science 305: 1163–1167.

    Article  CAS  Google Scholar 

  • Tatton L, Morley GM, Chopra R, Khwaja A . (2003). The src-selective kinase inhibitor PP1 also inhibits kit and bcr-abl tyrosine kinases. J Biol Chem 278: 4847–4853.

    Article  CAS  Google Scholar 

  • Turkson J, Bowman T, Garcia R, Caldenhoven E, De Groot RP, Jove R . (1998). Stat3 activation by src induces specific gene regulation and is required for cell transformation. Mol Cell Biol 18: 2545–2552.

    Article  CAS  Google Scholar 

  • Wiley HS, Burke PM . (2001). Regulation of receptor tyrosine kinase signaling by endocytic trafficking. Traffic 2: 12–18.

    Article  CAS  Google Scholar 

  • Yang S, Park K, Turkson J, Arteaga CL . (2008). Ligand-independent phosphorylation of Y869 (Y845) links mutant EGFR signaling to stat-mediated gene expression. Exp Cell Res 314: 413–419.

    Article  CAS  Google Scholar 

  • Yang S, Qu S, Perez-Tores M, Sawai A, Rosen N, Solit DB et al. (2006). Association with HSP90 inhibits cbl-mediated down-regulation of mutant epidermal growth factor receptors. Cancer Res 66: 6990–6997.

    Article  CAS  Google Scholar 

  • Yun CH, Boggon TJ, Li Y, Woo MS, Greulich H, Meyerson M et al. (2007). Structures of lung cancer-derived EGFR mutants and inhibitor complexes: mechanism of activation and insights into differential inhibitor sensitivity. Cancer Cell 11: 217–227.

    Article  CAS  Google Scholar 

  • Zhang J, Kalyankrishna S, Wislez M, Thilaganathan N, Saigal B, Wei W et al. (2007). SRC-family kinases are activated in non-small cell lung cancer and promote the survival of epidermal growth factor receptor-dependent cell lines. Am J Pathol 170: 366–376.

    Article  CAS  Google Scholar 

  • Zhang X, Gureasko J, Shen K, Cole PA, Kuriyan J . (2006). An allosteric mechanism for activation of the kinase domain of epidermal growth factor receptor. Cell 125: 1137–1149.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank members of the Band laboratories for helpful suggestions and discussion. This work was supported by: the NIH grants CA87986, CA105489, CA99900, CA116552 and CA99163 to HB, and CA94143, CA96844 and CA81076 to VB; Department of Defense Breast Cancer Research Grants W81XVVH-08-1-0617 (HB) and DAMD17-02-1-0508 (VB); the NCI Cancer Center of Nanotechnology Excellence Grant NCI 1U54 CA119341-01 (HB and VB); the NCI Breast Cancer SPORE and AVON Breast Cancer Fund at Robert H Lurie Cancer Center, Northwestern University; the H Foundation, Chicago, IL, USA; the Jean Ruggles-Romoser Chair of Cancer Research (HB) and the Duckworth Family Chair of Breast Cancer Research (VB); and the Malkin Scholarship (BMC). UNMC-Eppley Cancer Center is supported by an NCI Cancer Center Core Grant.

Author information

Author notes

  1. M Dimri

    Present address: 6Current address: Evanston Northwestern Healthcare Research Institute, Evanston, IL, USA.,

  2. A L Reddi

    Present address: 7Current address: Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.,

  3. G Chen

    Present address: 8Current address: Coskata Energy, Warrenville, IL, USA.,

  4. B M Chung: BMC is a Ph.D. degree candidate of the Interdepartmental Biological Sciences Program of Weinberg College of Arts and Sciences, Northwestern University, Evanston, IL, USA.

  5. B M Chung, M Dimri, M George, A L Reddi, G Chen, V Band and H Band: The work presented here was initiated while the principal investigators were at: 9Evanston Northwestern Healthcare Research Institute, Department of Medicine, Feinberg School of Medicine, Department of Biochemistry, Molecular Biology and Cell Biology, Weinberg College of Arts and Robert H. Lurie Comprehensive Cancer Center, Northwestern University, Evanston, IL, USA.

Authors and Affiliations

  1. Eppley Institute for Cancer and Allied Diseases, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE, USA

    B M Chung, M George & H Band

  2. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE, USA

    H Band

  3. Department of Genetics, Cell Biology and Anatomy, College of Medicine, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE, USA

    V Band & H Band

  4. UNMC-Eppley Cancer Center, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE, USA

    V Band & H Band

Authors
  1. B M Chung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M Dimri
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M George
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A L Reddi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. V Band
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H Band
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to H Band.

Additional information

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chung, B., Dimri, M., George, M. et al. The role of cooperativity with Src in oncogenic transformation mediated by non-small cell lung cancer-associated EGF receptor mutants. Oncogene 28, 1821–1832 (2009). https://doi.org/10.1038/onc.2009.31

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2009.31

Keywords

This article is cited by

Search

Advanced search

Quick links