Skip to main content

Advertisement

Log in

Dual targeting of EGFR and HER-2 in colon cancer cell lines

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

A number of studies have revealed that coexpression of EGFR and HER-2 has been found in a subset of colon cancers and may cooperatively promote tumor cell growth and survival. In the present work, two tyrosine kinase inhibitors, gefitinib and lapatinib, together with trastuzumab, raised a monoclonal antibody against HER-2 were evaluated in two colon cancer cell lines, DLD-1 and Caco-2. The aim of the study was to investigate their effect on tumor cell proliferation and apoptosis.

Methods

Cell proliferation was assessed using the MTT assay and apoptosis was evaluated by DNA fragmentation and the Annexin V binding assay. EGFR and HER-2 protein and mRNA levels were evaluated by immunoblotting and quantitative RT-PCR, respectively.

Results

Treatment of cells with each agent alone resulted in inhibition of cell proliferation after 48 h in a dose-dependent manner except for trastuzumab, which did not alter cell proliferation of DLD-1. Apoptosis increased in DLD-1 cells, after 24 h treatment with gefitinib. None of the tested agents altered apoptosis in Caco-2 cells. HER-2 and EGFR protein levels did not follow the changes of mRNA levels after treatment with the tested agents.

Conclusions

Τhe inhibitory effect of these agents on cell proliferation and the induction of apoptosis differ for the two colon cancer cell lines under consideration. Further studies are necessary to investigate the way they exert their antitumor effect.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Marmor MD, Skaria KB, Yarden Y (2004) Signal transduction and oncogenesis by ErbB/HER receptors. Int J Radiat Oncol Biol Phys 58:903–913

    PubMed  CAS  Google Scholar 

  2. Lee JC, Wang ST, Chow NH et al (2002) Investigation of the prognostic value of co expressed erbB family members for the survival of colorectal cancer patients after curative surgery. Eur J Cancer 38:1065–1071

    Article  PubMed  CAS  Google Scholar 

  3. Spano J, Fagard R, Soria JC et al (2005) Epidermal growth factor receptor signaling in colorectal cancer: preclinical data and therapeutic perspectives. Ann Oncol 16:189–194

    Article  PubMed  CAS  Google Scholar 

  4. Wakeling AE (2005) Inhibitors of growth factor signaling. Endoc Relat Cancer 12:S183–S187

    Article  CAS  Google Scholar 

  5. Jones HE, Gee JMW, Taylor KM et al (2005) Development of strategies for the use of anti-growth factor treatments. Endoc Relat Cancer 12:S173–S182

    Article  CAS  Google Scholar 

  6. Arora A, Scholar EM (2005) Role of tyrosine kinase inhibitors in cancer therapy. J Pharm Exp Ther 315:971–979

    Article  CAS  Google Scholar 

  7. Dancey J, Sausville E (2003) Issues and progress with protein kinase inhibitors for cancer treatment. Nat Rev Drug Discov 2:296–313

    Article  PubMed  CAS  Google Scholar 

  8. Alekshun T, Garrett C (2005) Targeted therapies in the treatment of colorectal cancers. Cancer control 12:105–110

    PubMed  Google Scholar 

  9. Saltz LB, Meropol NJ, Loehrer SRPJ et al (2004) Phase II trial of cetuximab in patients with refractory colorectal cancer that expresses the epidermal growth factor receptor. J Clin Oncol 22:1201–1208

    Article  PubMed  CAS  Google Scholar 

  10. Xu JM, Azzariti A, Colucci G et al (2003) The effect of gefitinib (Iressa, ZD1839) in combination with oxaliplatin is schedule-dependent in colon cancer cell lines. Cancer Chemother Pharmacol 52:442–448

    Article  PubMed  Google Scholar 

  11. Prewett MC, Hooper AT, Bassi R et al (2002) Enhanced antitumor activity of anti-epidermal growth factor receptor monoclonal antibody IMC-C225 in combination with irinotecan (CPT 11) against human colorectal tumor xenografts. Clin Cancer Res 8:994–1003

    PubMed  CAS  Google Scholar 

  12. Goldstein NI, Prewett M, Zuklys K et al (1995) Biological efficacy of a chimeric antibody to the epidermal growth factor receptor in a human tumor xenograft model. Clin Cancer Res 1:1311–1318

    PubMed  CAS  Google Scholar 

  13. Mann M, Sheng H, Shao J et al (2001) Targeting cyclooxygenase 2 and HER-2/neu pathways inhibits colorectal carcinoma growth. Gastroenterology 120:1713–1719

    Article  PubMed  CAS  Google Scholar 

  14. Roskoski R (2004) The ErbB/HER receptor protein-tyrosine kinases and cancer. Biochem Biophys Res Commun 319:1–11

    Article  PubMed  CAS  Google Scholar 

  15. Satoh H, Ishikawa H, Nakayama M et al (2004) Cell growth after withdrawal of gefitinib (‘‘Iressa’’, ZD1839), in human lung cancer cells. Oncol Rep 12:615–619

    PubMed  CAS  Google Scholar 

  16. Friedmann B, Caplin M, Hartley J et al (2004) Modulation of DNA repair in vitro after treatment with chemotherapeutic agents by the epidermal growth factor receptor inhibitor gefitinib (ZD1839). Clin Cancer Res 10:6476–6486

    Article  PubMed  CAS  Google Scholar 

  17. Vicentini C, Festuccia C, Gravina GL et al (2003) Prostate cancer cell proliferation is strongly reduced by the epidermal growth factor receptor tyrosine kinase inhibitor ZD1839 in vitro on human cell lines and primary cultures. J Cancer Res Clin Oncol 129:165–174

    PubMed  CAS  Google Scholar 

  18. Cunningham MP, Thomas H, Fan Z et al (2006) Responses of human colorectal tumor cells to treatment with the anti-epidermal growth factor receptor monoclonal antibody ICR62 used alone and in combination with the EGFR tyrosine kinase inhibitor gefitinib. Cancer Res 66:7708–7715

    Article  PubMed  CAS  Google Scholar 

  19. Spector N, Raefsky E, Hurwitz H et al (2003) Safety, clinical efficacy and biological assessments from EGF1004: a randomized phase IB study of GW572016 for patients with metastatic carcinomas expressing EGFR or ErbB2. Proc Am Soc Clin Oncol 22:193

    Google Scholar 

  20. Chu I, Blackwell K, Chen S et al (2005) The Dual ErbB1/ErbB2 Inhibitor, Lapatinib (LAPATINIB), cooperates with tamoxifen to inhibit both cell proliferation- and estrogen-dependent gene expression in antiestrogen-resistant breast cancer. Cancer Res 65:18–25

    PubMed  CAS  Google Scholar 

  21. Gregory C, Whang YE, McCall W et al (2005) Heregulin-induced activation of HER2 and HER3 increases androgen receptor transactivation and CWR-R1 human recurrent prostate cancer cell growth. Clin Cancer Res 11:1704–1712

    Article  PubMed  CAS  Google Scholar 

  22. Stulík J, Hernychová L, Porkertová S et al (2001) Proteome study of colorectal carcinogenesis. Electrophoresis 22:3019–3025

    Article  PubMed  Google Scholar 

  23. Kuwada SK, Scaife CL, Kuang J et al (2004) Effects of trastuzumab on epidermal growth factor receptor-dependent and–independent human colon cancer cells. Int J Cancer 109:291–301

    Article  PubMed  CAS  Google Scholar 

  24. Rusnak DW, Lackey K, Affleck K et al (2001) The effects of the novel, reversible epidermal growth factor receptor/ErbB-2 tyrosine kinase inhibitor, GW2016, on the growth of human normal and tumor-derived cell lines in vitro and in vivo. Mol Cancer Ther 1:85–94

    PubMed  CAS  Google Scholar 

  25. Kuwahara Y, Hosoi H, Osone S et al (2004) Antitumor activity of gefitinib in malignant rhabdoid tumor cells in vitro and in vivo. Clin Cancer Res 10:5940–5948

    Article  PubMed  CAS  Google Scholar 

  26. Zhou Y, Li S, Hu YP et al (2006) Blockade of EGFR and ErbB2 by the novel dual EGFR and ErbB2 tyrosine kinase inhibitor GW572016 sensitizes human colon carcinoma GEO cells to apoptosis. Cancer Res 66:404–411

    Article  PubMed  CAS  Google Scholar 

  27. Giannopoulou E, Papadimitriou E (2003) Amifostine has antiangiogenic properties in vitro by changing the redox status of human endothelial cells. Free Rad Res 37:1191–1199

    Article  CAS  Google Scholar 

  28. Normanno N, Campiglio M, De Luca A et al (2002) Cooperative inhibitory effect of ZD1839 (Iressa) in combination with trastuzumab (Herceptin) on human breast cancer cell growth. Ann Oncol 13:65–72

    Article  PubMed  CAS  Google Scholar 

  29. Loeffler-Ragg J, Skvortsov S, Sarg B et al (2005) Gefitinib-responsive EGFR-positive colorectal cancers have different proteome profiles from non-responsive cell lines. Eur J Cancer 41:2338–2346

    Article  PubMed  CAS  Google Scholar 

  30. Duneau JP, Vegh AP, Sturgis JN (2007) A dimerization hierarchy in the transmembrane domains of the HER receptor family. Biochemistry 46:2010–2019

    Article  PubMed  CAS  Google Scholar 

  31. Rabindran SK (2005) Antitumor activity of HER-2 inhibitors. Cancer Lett 227:9–23

    Article  PubMed  CAS  Google Scholar 

  32. Ferrer-Soler L, Vazquez-Martin A, Brunet J (2007) An update of the mechanisms of resistance to EGFR-tyrosine kinase inhibitors in breast cancer: gefitinib (Iressa)–induced changes in the expression and nucleo-cytoplasmic trafficking of HER-ligands (Review). Int J Mol Med 20:3–10

    PubMed  CAS  Google Scholar 

  33. Li X, Leu S, Cheong A et al (2004) Akt2, phosphatidylinositol 3-kinase, and PTEN are in lipid rafts of intestinal cells: role in absorption and differentiation. Gastroenterology 126:122–135

    Article  PubMed  CAS  Google Scholar 

  34. Fischel JL, Formento P, Milano G (2005) Epidermal growth factor receptor doubles targeting by a tyrosine kinase inhibitor (Iressa) and a monoclonal antibody (Cetuximab). Impact on cell growth and molecular factors. Br J Cancer 92:1063–1068

    Article  PubMed  CAS  Google Scholar 

  35. Xia W, Mullin RJ, Keith BR et al (2002) Anti-tumor activity of GW572016: a dual tyrosine kinase inhibitor blocks EGF activation of EGFR/erbB2 and downstream Erk1/2 and AKT pathways. Oncogene 21:6255–6263

    Article  PubMed  CAS  Google Scholar 

  36. Okochi-Takada E, Nakazawa K, Wakabayashi M (2006) Silencing of the UCHL1 gene in human colorectal and ovarian cancers. Int J Cancer 119:1338–1344

    Article  PubMed  CAS  Google Scholar 

  37. Nahta R, Yu D, Hung MC et al (2006) Mechanisms of Disease: understanding resistance to HER2-targeted therapy in human breast cancer. Nat Clin Pract Oncol 3:269–280

    Article  PubMed  CAS  Google Scholar 

  38. Lane HA, Beuvink I, Motoyama AB et al (2000) ErbB2 potentiates breast tumor proliferation through modulation of p27Kip1-Cdk2 complex formation: receptor overexpression does not determine growth dependency. Mol Cell Biol 20:3210–3223

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Efstathia Giannopoulou and Anna Antonacopoulou were supported by Hellenic Cooperative Oncology Group (HECOG).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Haralabos P. Kalofonos.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Giannopoulou, E., Antonacopoulou, A., Floratou, K. et al. Dual targeting of EGFR and HER-2 in colon cancer cell lines. Cancer Chemother Pharmacol 63, 973–981 (2009). https://doi.org/10.1007/s00280-008-0820-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-008-0820-9

Keywords

Navigation