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The sensitivity of the in vitro cytokinesis-blocked micronucleus assay in lymphocytes for different and combined radiation qualities

Die Sensitivität des In-vitro-Zytokinesis-Block-Mikronukleus-Assays für verschiedene Strahlenqualitäten und deren Kombination

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Abstract

Purpose

The dose-response relationship and the relative biological effectiveness (RBE) for the induction of micronuclei in lymphocytes was analyzed after irradiation in vitro with a 6-MeV neutron beam that was followed by 240-kV X-rays. The dose range of the combined exposure comprised 1 to 3 Gy. For reference, the dose-effect relationships found after X-ray (0.5 to 5 Gy)-and neutron (0.5 to 4 Gy) exposure applied separately are presented. The possibility of an interaction between the 2 radiation qualities is investigated by the method of isobole calculation termed “envelope of additivity”.

Methods

Micronuclei were analyzed in PHA-stimulated, cytokinesis-blocked human lymphocytes.

Results

The dose-response relationships for the micronucleus frequencies induced by the neutron irradiation, as well as by the mixed exposure, were linear. A saturation effect was indicated after neutron doses higher than 3 Gy. After low LET exposure the dose-response curves were describable by a linear-quadratic model. For neutron-induced micronucleus frequencies, RBE-values of 2 to 3 and for the combined exposure RBE values of 1.5 to 2 were calculated for a range of effect of 0.5 to 1.5 micronuclei/binucleated lymphocyte. No indication was found for an interaction between the damage induced by X-rays and that produced by neutrons under our experimental conditions.

Conclusions

These studies demonstrate a clear dependence of micronucleus induction on radiation quality and empha-size the usefulness of the micronucleus assay in biological dosimetry, also in cases in which high LET radiation or a mixed beam is involved as the radiation source.

Zusammenfassung

Fragestellung

Die Mikronukleusinduktion in Lymphozyten wurde nach In-vitro-Bestrahlung mit 6-MeV-Neutronen (0,5 bis 4 Gy), 240-kV-Röntgenstrahlung (0,5 bis 5 Gy) bzw. einer Kombination dieser Strahlenqualitäten (1 bis 3 Gy Gesamtdosis) untersucht. Anhand der Dosis-Wirkungs-Beziehungen für die einzelne und kombinierte Anwendung beider Strahlenarten wurde die relative biologische Wirksamkeit (RBW) für Neutronen bzw. für die Kombination von Neutronen und Röntgenstrahlen ermittelt. Mit Hilfe einer Isobolenkalkulation (“envelope of additivity”) wurde die Möglichkeit einer Interaktion zwischen den beiden Strahlenqualitäten analysiert.

Methode

Die Auswertung der Mikronuklei erfolgte in PHA-stimulierten humanen Lymphozyten, deren Zytokinese durch Cytochalasin B inhibiert worden war.

Ergebnisse

Für die Neutronen-wie auch für die kombinierte Bestrahlung ergaben sich jeweils lineare Dosis-Wirkungs-Beziehungen. Ein Sättigungseffekt stellte sich für Neutronendosen oberhalb von 3 Gy ein. Nach Niedrig-LET-Bestrahlung folgte die Dosis-Wirkungs-Beziehung dem linear-quadratischen Modell. Ein Vergleich der für die Induktion von 0,5 bis 1,5 Mikronuklei pro binukleärer Zelle isoeffektiven Dosen ergab für die Bestrahlung mit Neutronen RBW-Werte von 2 bis 3 und für die kombinierte Bestrahlung Werte von 1,5 bis 2 unter den von uns gewählten experimentellen Bedingungen. Es konnte kein Hinweis auf eine Interaktion zwischen den durch die Röntgen-bzw. Neutronenstrahlung induzierten Schäden gewonnen werden.

Schlußfolgerungen

Die Untersuchungen zeigen eine deutliche Abhängigkeit der Mikronukleusbildung von der Strahlenqualität. Die Ergebnisse verdeutlichen, daß der Einsatz des Mikronukleusassays in der biologischen Dosimetrie auch dann sinnvoll ist, wenn als Strahlenquelle eine Hoch-LET-Strahlung bzw. eine Mischung aus Hoch-und Niedrig-LET-Strahlen vorliegt.

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References

  1. Bauchinger M, Schmid E, Rimpl G, et al. Chromosome aberrations in human lymphocytes after irradiation with 15.0-MeV neutrons in vitro. I. Dose-response relation and RBE. Mutation Res 1975;27:103–9

    Article  PubMed  CAS  Google Scholar 

  2. Beuningen D van, Streffer C, Bertholdt G. Mikronukleusbildung im Vergleich zur Überlebensrate von menschlichen Melanomzellen nach Röntgen-, Neutronenbestrahlung und Hyperthermie. Strahlentherapie 1981;157:600–6.

    PubMed  Google Scholar 

  3. Bilbao A, Prosser JS, Edwards AA, et al. The induction of micronuclei in human lymphocytes by in vitro irradiation with alpha particles from plutonium-239. Int J Radiat Biol 1989;56:287–92.

    Article  PubMed  CAS  Google Scholar 

  4. Böcker W, Streffer C, Müller W-U, et al. Automated scoring of micronuclei in binucleated human lymphocytes. Int J Radiat Res 1996; 70:529–37.

    Google Scholar 

  5. Brooks AL, Newton GJ, Shyr LJ, et al. The combined effects of alpha particles and X-rays on cell killing and micronuclei induction in lung epithelial cells. Int J Radiat Biol 1990;58:799–811.

    Article  PubMed  CAS  Google Scholar 

  6. Evans HJ. Mutation cytogenetics: past, present and future. Mutation Res 1988;204:355–63.

    Article  PubMed  CAS  Google Scholar 

  7. Fuhrmann C, Streffer C, Müller W-U, et al. Micronucleus asay prediction and application optimized by cytochalasin B-induced binucleated tumor cells. Strahlenther Onkol 1992;10:603–9.

    Google Scholar 

  8. Gantenberg H-W, Wuttke K, Streffer C, et al. Micronuclei in human lymphocytes irradiated in vitro or in vivo. Radiat Res 1991;128:276–81.

    Article  PubMed  CAS  Google Scholar 

  9. Huber R, Schraube H, Bauchinger M. In vitro induction of micronuclei in human lymphocytes by fission neutrons. Programme and Abstract Book, 24th Annual Meeting of the European Society for Radiation Biology, Erfurt, Germany, 1992:175.

  10. Huber R, Schraube H, Nahrstedt U, et al. Dose-response relationships of micronuclei in human lymphocytes induced by fission neutrons and by low LET radiations. Mutation Res 1994;306:135–41.

    PubMed  CAS  Google Scholar 

  11. Kim SH, Cho CK, Kim TH, et al. Frequency of micronuclei in lymphocytes following gamma and fast-neutron irradiations. Anticancer Res 1993;13:1587–92.

    PubMed  CAS  Google Scholar 

  12. Littlefield LG, Sayer AM, Frome EL. Comparison of dose-response parameters for radiation-induced acentric fragments and micronuclei observed in cytokinesis-arrested lymphocytes. Mutagenesis 1989;4:265–70.

    Article  PubMed  CAS  Google Scholar 

  13. Lloyd DC, Purrott RJ, Dolphin GW, et al. Chromosome aberrations induced in human lymphocytes by neutron irradiation. Int J Radiat Biol 1976;29:169–82.

    Article  CAS  Google Scholar 

  14. Molls M, Streffer C, Zamboglou N. Micronucleus formation in preimplanted mouse embryos cultured in vitro after irradiation with X-rays and neutrons. Int J Radiat Biol 1981;39:307–14.

    Article  CAS  Google Scholar 

  15. Müller W-U, Streffer C. Biological indicators for radiation damage. Int J Radiat Biol 1991;59:863–73.

    Article  PubMed  Google Scholar 

  16. Müller W-U, Streffer C. Micronucleus assays. In: Obe G, ed. Advances in mutagenesis research Vol. 4, Berlin: Springer, 1994:1–133.

    Google Scholar 

  17. Murthy MSS, Madhvanath U, Subrahmanyam P, et al. Synergistic effect of simultaneous exposure to Co-60 gamma rays and Po-210 alpha rays in diploid yeast. Radiat Res 1975;63:185–90.

    Article  PubMed  CAS  Google Scholar 

  18. Ono K, Nagata Y, Akuta K, et al. Frequency of micronuclei in hepatocytes following X and fast neutron irradiations — An analysis by a linearquadratic model. Radiat Res 1990;123:345–47.

    Article  PubMed  CAS  Google Scholar 

  19. Pampfer S, Müller W-U, Streffer C. Preimplantation growth delay and micronucleus formation after in vivo exposure of mouse zygotes to fast neutrons. Radiat Res 1992;129:88–95.

    Article  PubMed  CAS  Google Scholar 

  20. Prosser JS, Moquet JE, Lloyd DC, et al. Radiation induction of micronuclei in human lymphocytes. Mutation Res 1988;199:37–45.

    PubMed  CAS  Google Scholar 

  21. Rassow J. Medical cyclotron facilities — Useful tools for clinical therapy, diagnosis and analysis. Proc Int Symp Appl Technol Ion Radiat 1982;1: 47–100.

    Google Scholar 

  22. Revell SH. Relationships between chromosome damage and cell death. In: Liss AR, ed. Radiation-induced chromosome damage in man. New York 1983:215–33.

  23. Steel GG, Peckham MJ. Expoitable mechanisms in combined radiotherapy-chemotherapy: the concept of additivity. Int J Radiat Oncol Biol Phys 1979;5:85–91.

    PubMed  CAS  Google Scholar 

  24. Straume T, Lucas JN, Tucker JD, et al. Biodosimetry for a radiation worker using multiple assays. Health Phys 1992;62:122–30.

    Article  PubMed  CAS  Google Scholar 

  25. Streffer C, Müller W-U. Radiation risk from combined exposures to ionizing radiations and chemicals. II. Terminology and mode of interaction. Adv Radiat Biol 1984;11:175–80.

    Google Scholar 

  26. Vral A, Verhaegen F, Thierens H, et al. Micronuclei induced by fast neutrons versus 60Co gamma-rays in human peripheral blood lymphocytes. Int J Radiat Biol 1994;65:321–28.

    Article  PubMed  CAS  Google Scholar 

  27. Wuttke K, Streffer C, Müller W-U, Radiation induced micronuclei in subpopulations of human lymphocytes. Mutation Res 1993;286:181–88.

    PubMed  CAS  Google Scholar 

  28. Wuttke K, Müller W-U, Streffer C. Micronucleus expression in human lymphocytes after in vitro exposure to X-rays, neutrons or neutrons followed by X-rays. Programme and Abstract Book, 26th Annual Meeting of the European Society for Radiation Biology, Amsterdam, 1994:137.

  29. Wuttke K, Streffer C, Müller W-U, et al. Micronuclei in lymphocytes of children from the vicinity of Chernobyl before and after I-131 therapy for thyroid cancer. Int J Radiat Biol 1996;69:259–68.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Institut für Medizinische Strahlenbiologie, Universitätsklinikum Essen, Hufelandstraße 55, D-45122, Essen, Germany

    Kerstin Wuttke, Wolfgang-Ulrich Müller & Christian Streffer

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  1. Kerstin Wuttke
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  2. Wolfgang-Ulrich Müller
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  3. Christian Streffer
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Wuttke, K., Müller, WU. & Streffer, C. The sensitivity of the in vitro cytokinesis-blocked micronucleus assay in lymphocytes for different and combined radiation qualities. Strahlenther Onkol 174, 262–268 (1998). https://doi.org/10.1007/BF03038719

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  • DOI: https://doi.org/10.1007/BF03038719

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