Differences in radiosensitivity between three HER2 overexpressing cell lines

  • Ann-Charlott Steffen
  • Lovisa Göstring
  • Vladimir Tolmachev
  • Stig Palm
  • Bo Stenerlöw
  • Jörgen Carlsson
Original Article

Abstract

Purpose

HER2 is a potential target for radionuclide therapy, especially when HER2 overexpressing breast cancer cells are resistant to Herceptin® treatment. Therefore, it is of interest to analyse whether HER2 overexpressing tumour cells have different inherent radiosensitivity.

Methods

The radiosensitivity of three often used HER2 overexpressing cell lines, SKOV-3, SKBR-3 and BT-474, was analysed. The cells were exposed to conventional photon irradiation, low linear energy transfer (LET), to characterise their inherent radiosensitivity. The analysis was made with clonogenic survival and growth extrapolation assays. The cells were also exposed to alpha particles, high LET, from 211At decays using the HER2-binding affibody molecule 211At-(ZHER2:4)2 as targeting agent. Assays for studies of internalisation of the affibody molecule were applied.

Results

SKOV-3 cells were most radioresistant, SKBR-3 cells were intermediate and BT-474 cells were most sensitive as measured with the clonogenic and growth extrapolation assays after photon irradiation. The HER2 dependent cellular uptake of 211At was qualitatively similar for all three cell lines. However, the sensitivity to the alpha particles from 211At differed; SKOV-3 was most resistant, SKBR-3 intermediate and BT-474 most sensitive. These differences were unexpected because it is assumed that all types of cells should have similar sensitivity to high-LET radiation. The sensitivity to alpha particle exposure correlated with internalisation of the affibody molecule and with size of the cell nucleus.

Conclusion

There can be differences in radiosensitivity, which, if they also exist between patient breast cancer cells, are important to consider for both conventional radiotherapy and for HER2-targeted radionuclide therapy.

Keywords

ErbB2 HER2 High LET Nuclear medicine Radiation effects Tumour cells 

References

  1. 1.
    Adams GP, Weiner LM. Monoclonal antibody therapy of cancer. Nat Biotechnol 2005;23(9):1147–57.PubMedCrossRefGoogle Scholar
  2. 2.
    DeNardo SJ, DeNardo GL. Targeted radionuclide therapy for solid tumors: an overview. Int J Radiat Oncol Biol Phys 2006;66(2 Suppl):S89–95.PubMedGoogle Scholar
  3. 3.
    Goldenberg DM, Sharkey RM. Advances in cancer therapy with radiolabeled monoclonal antibodies. Q J Nucl Med Mol Imaging 2006;50(4):248–64.PubMedGoogle Scholar
  4. 4.
    O’Donoghue JA, Bardies M, Wheldon TE. Relationships between tumor size and curability for uniformly targeted therapy with beta-emitting radionuclides. J Nucl Med 1995;36(10):1902–9.PubMedGoogle Scholar
  5. 5.
    Carlsson J, Forssell AE, Hiet-ala SO, Stigbrand T, Tennvall J. Tumour therapy with radionuclides: assessment of progress and problems. Radiother Oncol 2003;66:107–17.PubMedCrossRefGoogle Scholar
  6. 6.
    Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 1987;235:177–82.PubMedCrossRefGoogle Scholar
  7. 7.
    Berchuck A, Kamel A, Whitaker R, Kerns B, Olt G, Kinney R, et al. Overexpression of HER-2/neu is associated with poor survival in advanced epithelial ovarian cancer. Cancer Res 1990;50:4087–91.PubMedGoogle Scholar
  8. 8.
    Gancberg D, Di Leo A, Cardoso F, Rouas G, Pedrocchi M, Paesmans M, et al. Comparison of HER-2 status between primary breast cancer and corresponding distant metastatic sites. Ann Oncol 2002;13:1036–43.PubMedCrossRefGoogle Scholar
  9. 9.
    Carlsson J, Nordgren H, Sjostrom J, Wester K, Villman K, Bengtsson NO, et al. HER2 expression in breast cancer primary tumours and corresponding metastases. Original data and literature review. Br J Cancer 2004;90:2344–8.PubMedGoogle Scholar
  10. 10.
    Steel GG. Basic clinical radiobiology. London: Hodder Education; 2002.Google Scholar
  11. 11.
    Hall EJ, Giaccia AJ. Radiobiology for the radiologist. Philadelphia: Lippincott Williams & Wilkins; 2006.Google Scholar
  12. 12.
    Deacon J, Peckham MJ, Steel GG. The radioresponsiveness of human tumours and the initial slope of the cell survival curve. Radiother Oncol 1984;2:317–23.PubMedCrossRefGoogle Scholar
  13. 13.
    Deschavanne PJ, Fertil B. A review of human cell radiosensitivity in vitro. Int J Radiat Oncol Biol Phys 1996;34(1):251–66.PubMedGoogle Scholar
  14. 14.
    Karlsson KH, Stenerlow B. Focus formation of DNA repair proteins in normal and repair-deficient cells irradiated with high-LET ions. Radiat Res 2004;161:517–27.PubMedCrossRefGoogle Scholar
  15. 15.
    Johansson L, Nilsson K, Carlsson J, Larsson B, Jakobsson P. Radiation effects on cultured human lymphoid cells. Analysis using the growth extrapolation method. Acta Radiol Oncol 1981;20:51–9.PubMedCrossRefGoogle Scholar
  16. 16.
    Persson MI, Gedda L, Jensen HJ, Lundqvist H, Malmstrom PU, Tolmachev V. Astatinated trastuzumab, a putative agent for radionuclide immunotherapy of ErbB2-expressing tumours. Oncol Rep 2006;15(3):673–80.PubMedGoogle Scholar
  17. 17.
    Wikman M, Steffen AC, Gunneriusson E, Tolmachev V, Adams GP, Carlsson J, et al. Selection and characterization of HER2/neu-binding affibody ligands. Protein Eng DeS Selection (PEDS) 2004;17(5):455–62.CrossRefGoogle Scholar
  18. 18.
    Steffen AC, Wikman M, Tolmachev V, Adams GP, Nilsson F, Ståhl S, et al. In vitro characterization of a bivalent anti-HER2 affibody with potential for radionuclide based diagnostics. Cancer Biother Radiopharm 2005;20(3):239–48.PubMedCrossRefGoogle Scholar
  19. 19.
    Steffen AC, Orlova A, Wikman M, Nilsson F, Ståhl S, Adams GP, et al. Affibody mediated tumour targeting of HER-2 expressing xenografts in mice. Eur J Nucl Med Mol Imaging 2006;33(6):631–8.PubMedCrossRefGoogle Scholar
  20. 20.
    Orlova A, Magnusson M, Eriksson T, Nilsson M, Larsson B, Höiden-Guthenberg I, et al. Tumor imaging using a picomolar affinity HER-2 binding affibody molecule. Cancer Res 2006;66(8):4339–433.PubMedCrossRefGoogle Scholar
  21. 21.
    Tolmachev V, Orlova A, Pehrson R, Galli J, Baastorp B, Andersson K, et al. Radionuclide therapy of HER2-positive micro-xenografts using a 177-Lu-labeled HER2-specific Affibody molecule. Cancer Res 2007;67(6):2773–82.PubMedCrossRefGoogle Scholar
  22. 22.
    Hartman T. Tumour targeting with stable and radioactive nuclides: dosimetric aspects at the cellular level. Doctoral thesis, Uppsala University. Uppsala: Acta Universitatis Upsaliensis; 1999.Google Scholar
  23. 23.
    Palm S, Humm JL, Rundqvist R, Jacobsson L. Microdosimetry of astatine-211 single-cell irradiation: role of daughter polonium-211 diffusion. Med Phys 2004;31:218–25.PubMedCrossRefGoogle Scholar
  24. 24.
    Palm S. In vitro effects and microdosimetry of astatine-211 for tumour therapy. Doctoral thesis, University of Göteborg, Sahlgrenska University Hospital (ISBN 91-628-3841-5); 1999.Google Scholar
  25. 25.
    Nordberg E, Friedman M, Göstring L, Adams GP, Brismar H, Nilsson FY, et al. Cellular studies of binding, internalization and retention of a radiolabeled EGFR-binding affibody molecule. Nucl Med Biol 2007;34(6):609–18.PubMedCrossRefGoogle Scholar
  26. 26.
    Dikomey E, Brammer I. Relationship between cellular radiosensitivity and non-repaired double-strand breaks studied for different growth states, dose-rates and plating conditions in a normal human fibroblast line. Int J Radiat Biol 2000;76(6):773–81.PubMedCrossRefGoogle Scholar
  27. 27.
    Yaginuma Y, Westphal H. Abnormal structure and expression of the p53 gene in human ovarian carcinoma cell lines. Cancer Res 1992;52:4196–9.PubMedGoogle Scholar
  28. 28.
    Carlsson J, Hakansson E, Eriksson V, Grawe J, Wester K, Grusell E, et al. Early effects of low dose-rate radiation on cultured tumor cells. Cancer Biother Radiopharm 2003b;18:663–70.PubMedCrossRefGoogle Scholar
  29. 29.
    Lennartsson J, Carlsson J, Stenerlöw B. Targeting the epidermal growth factor receptor family in radionuclide therapy of tumors–signal transduction and DNA repair. Lett Drug Des Discov 2006;3:357–68.CrossRefGoogle Scholar
  30. 30.
    Petru E, Sevin BU, Gottlieb C. Radiosensitivity patterns of four human ovarian cancer cell lines in vitro. Gynecol Oncol 1997;64:490–2.PubMedCrossRefGoogle Scholar
  31. 31.
    Steren A, Sevin BU, Perras J, Angioli R, Nguyen H, Guerra L, et al. Taxol sensitizes human ovarian cancer cells to radiation. Gynecol Oncol 1993;48(2):252–8.PubMedCrossRefGoogle Scholar
  32. 32.
    Joiner MC, Marples B, Lambin P, Short SC, Turesson I. Low-dose hypersensitivity: current status and possible mechanisms. Int J Radiat Oncol Biol Phys 2001;49(2):379–89.PubMedCrossRefGoogle Scholar
  33. 33.
    Murray D, McEwan AJ. Radiobiology of systemic radiation therapy. Cancer Biother Radiopharm 2007;22(1):1–23.PubMedCrossRefGoogle Scholar
  34. 34.
    Chapman JD. Single-hit mechanism of tumour cell killing by radiation. Int J Radiat Biol 2003;79:71–81.PubMedCrossRefGoogle Scholar
  35. 35.
    Prise KM, Folkard M, Michael BD. A review of the bystander effect and its implications for low-dose exposure. Radiat Prot Dosim 2003;104(4):347–55.Google Scholar
  36. 36.
    Stenerlow B, Pettersson O, Essand M, Blomquist E, Carlsson J. Irregular variations in radiation sensitivity when the linear energy transfer is increased. Radiother Oncol 1995;36(2):133–42.PubMedCrossRefGoogle Scholar
  37. 37.
    Ekerljung L, Steffen AC, Carlsson J, Lennartsson J. Effects of HER2-binding affibody molecules on intracellular signaling pathways. Tumour Biol 2006;27(4):201–10.PubMedCrossRefGoogle Scholar
  38. 38.
    Zalutsky MR, Pozzi OR. Radioimmunotherapy with alpha-particle emitting radionuclides. Q J Nucl Med Mol Imaging 2004;48:289–96.PubMedGoogle Scholar
  39. 39.
    Smith-Jones PM, Solit DB, Akhurst T, Afroze F, Rosen N, Larson SM. Imaging the pharmacodynamics of HER2 degradation in response to Hsp90 inhibitors. Nat Biotechnol 2004;22(6):701–6.PubMedCrossRefGoogle Scholar
  40. 40.
    Smith-Jones PM, Solit D, Afroze F, Rosen N, Larson SM. Early tumor response to Hsp90 therapy using HER2 PET: comparison with 18F-FDG PET. J Nucl Med 2006;47(5):793–6.PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Ann-Charlott Steffen
    • 1
  • Lovisa Göstring
    • 2
  • Vladimir Tolmachev
    • 1
  • Stig Palm
    • 3
  • Bo Stenerlöw
    • 1
  • Jörgen Carlsson
    • 1
    • 4
  1. 1.Unit of Biomedical Radiation Sciences, Department of Oncology, Radiology and Clinical Immunology, Rudbeck LaboratoryUppsala UniversityUppsalaSweden
  2. 2.Affibody ABBrommaSweden
  3. 3.Department of Radiation PhysicsSahlgrenska Academy at Göteborg UniversityGöteborgSweden
  4. 4.Biomedical Radiation SciencesRudbeck LaboratoryUppsalaSweden

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