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DNA damage in subpopulations of human lymphocytes irradiated with doses in the range of 0-1 Gy of X-radiation

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EN
Abstrakty
EN
We compared three methods usually applied in biological dosimetry for estimation of radiation-induced DNA damage in human T and B lymphocytes: alkaline comet assay, micronucleus (MN) test and formation of histone gamma-H2AX foci. Human peripheral blood lymphocytes were fractionated using T cells and B cells isolation kits. Cells were irradiated with doses in the range of 0-1 Gy of X-rays. Induction of DNA damage was assessed by the standard alkaline comet assay, MN test and histone gammaH2AX foci immunofluorescence assay. Notwithstanding different end-points measured by the applied methods, all tests revealed a similar induction of DNA damage in B lymphocytes as compared with T lymphocytes. The results indicated that all three tests detect DNA damage with similar sensitivity, the lowest dose being approximately 0.3 Gy. The difference between irradiated and control cells was expressed as the ratio of the value obtained for irradiated cells (1 Gy) to that for control cells. The highest ratio was obtained for formation of gammaH2AX foci and was 6.2 for T and 13.8 for B lymphocytes, whereas those for comet assay and micronucleus test were 3.5; 3.6 and 5.6; 4.8, respectively.
Czasopismo
Rocznik
Strony
145--149
Opis fizyczny
Bibliogr. 17 poz., rys.
Twórcy
autor
autor
autor
autor
autor
  • Department of Radiobiology and Health Protection, Chemistry and Technology, 16 Dorodna Str., 03-195 Warsaw, Poland, Tel.: +48 22 504 11 74, Fax: +48 22 504 13 41, marylaw@ichtj.waw.pl
Bibliografia
  • 1. Almássy Z, Krepinsky AB, Bianco A, Köteles GJ (1987) The present state and perspectives of micronucleus assay in radiation protection. A review. Appl Radiat Isot 38:241–249
  • 2. Banath JP, MacPhail SH, Olive PL (2004) Radiation sensitivity, H2AX phosphorylation, and kinetics of repair of DNA strand breaks in irradiated cervical cancer cell lines. Cancer Res 64:7144–7149
  • 3. Cowell IG, Durkacz BW, Tilby MJ (2005) Sensitization of breast carcinoma cells to ionizing radiation by small molecule inhibitors of DNA-dependent protein kinase and ataxia telangiectasia mutated. Biochem Pharmacol 71:13–20
  • 4. Fenech M, Morley AA (1986) Cytokinesis-block micronucleus method in human lymphocytes: effect of in vivo ageing and low dose X-irradiation. Mutat Res 161:193–198
  • 5. Högstedt B, Karlsson A, Holmén A (1988) Frequencies of micronuclei in human peripheral blood lymphocytes stimulated in vitro by phytohemagglutinin and pokeweed mitogen. Hereditas 109:53–55
  • 6. Krol M, Lankoff A, Buraczewska I, Derezińska E, Wójcik A (2007) Radiation-induced micronucleus frequencies in female peripheral blood lymphocytes collected during the first and second half of the menstrual cycle. Radiat Prot Dosim 123:483–489
  • 7. Kruszewski M, Wojewódzka M, Iwaneńko T, Collins AR, Szumiel I (1998) Application of the comet assay for monitoring DNA damage in workers exposed to chronic low-dose irradiation. II. Base damage. Mutat Res 416:37–57
  • 8. Miller K (1983) The stimulation of human B and T lymphocytes by various lectins. Immunobiology 165:132–146
  • 9. Pouget JP, Ravanat JL, Douki T, Richard MJ, Cadet J (1999) Measurement of DNA base damage in cells exposed to low doses of gamma-radiation: comparison between the HPLC-EC and comet assays. Int J Radiat Biol 75:51–58
  • 10. Rao BS, Natarajan AT (2001) Retrospective biological dosimetry of absorbed radiation. Radiat Prot Dosim 95:17–23
  • 11. Rothkamm K, Lobrich M (2003) Evidence for a lack of DNA double-strand break repair in human cells exposed to very low X-ray doses. Proc Natl Acad Sci USA 100:5057–5062
  • 12. Schmitz A, Bayer J, Dechamps N, Goldin L, Thomas G (2007) Heritability of susceptibility to ionizing radiationinduced apoptosis of human lymphocyte subpopulations. Int J Radiat Oncol Biol Phys 68:1169–1177
  • 13. Schmitz A, Bayer J, Déchamps N, Thomas G (2003) Intrinsic susceptibility to radiation-induced apoptosis of human lymphocyte subpopulations. Int J Radiat Oncol Biol Phys 57:769–778
  • 14. Sedelnikova OA, Rogakou EP, Panyutin IG, Bonner WM (2002) Quantitative detection of (125)IdU-induced DNA double-strand breaks with gamma-H2AX antibody. Radiat Res 158:486–492
  • 15. Szumiel I (2007) Analysis of DNA double-strand breaks by means of γ-H2AX foci. In: Obe G, Vijayalaxmi G (eds) Chromosomal alterations: methods, results and importance in human health. Springer-Verlag, Berlin-Heidelberg, pp 145–160
  • 16. Wojewódzka M, Kruszewski M, Iwaneńko T, Collins AR, Szumiel I (1998) Application of the comet assay for monitoring DNA damage in workers exposed to chronic low-dose irradiation. I. Strand breakage. Mutat Res 416:21–35
  • 17. Wuttke K, Streffer C, Muller WU (1993) Radiation induced micronuclei in subpopulations of human lymphocytes. Mutat Res 286:181–188
Typ dokumentu
Bibliografia
Identyfikator YADDA
bwmeta1.element.baztech-article-BUJ6-0023-0034
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