Artículo

Bracalente, C.; Ibañez, I.L.; Molinari, B.; Palmieri, M.; Kreiner, A.; Valda, A.; Davidson, J.; Durán, H. "Induction and persistence of large γh2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells" (2013) International Journal of Radiation Oncology Biology Physics. 87(4):785-794
Estamos trabajando para incorporar este artículo al repositorio
Consulte el artículo en la página del editor
Consulte la política de Acceso Abierto del editor

Abstract:

Purpose: To evaluate the cell response to DNA double-strand breaks induced by low and high linear energy transfer (LET) radiations when the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), an essential protein of the nonhomologous end-joining repair pathway, lacks kinase activity. Methods and Materials: CHO10B2, a Chinese hamster ovary cell line, and its derived radiosensitive mutant cell line, irs-20, lacking DNA-PKcs activity, were evaluated after 0 to 3 Gy of γ-rays, plateau and Bragg peak protons, and lithium beams by clonogenic assay, and as a measurement of double-strand breaks, phosphorylated H2AX (γH2AX) foci number and size were quantified by immunocytofluorescence. Results: Irs-20 exhibited greater radiosensitivity and a higher amount of γH2AX foci than CHO10B2 at 6 hours after irradiation for all types of radiations. Remarkably, CHO10B2 and irs-20 maintained their difference in radiosensitivity after high-LET radiation. Six hours after low-LET radiations, irs-20 did not reach basal levels of γH2AX at high doses, whereas CHO10B2 recovered basal levels for all doses. After high-LET radiation, only CHO10B2 exhibited a reduction in γH2AX foci, but it never reached basal levels. Persistent foci in irs-20 confirmed a repair deficiency. Interestingly, after 30 minutes of high-LET radiation both cell lines exhibited large foci (size >0.9 μm2) related to the damage nature, whereas at 6 hours irs-20 showed a higher amount of large foci than CHO10B2, with a 7-fold increase at 3 Gy, that could also be associated to radiosensitivity. Conclusions: We demonstrated, for the first time, an association between deficient DNA-PKcs activity and not only high levels of H2AX phosphorylation but also persistence and size increase of γH2AX foci after high-LET irradiation. © 2013 Elsevier Inc.

Registro:

Documento: Artículo
Título:Induction and persistence of large γh2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells
Autor:Bracalente, C.; Ibañez, I.L.; Molinari, B.; Palmieri, M.; Kreiner, A.; Valda, A.; Davidson, J.; Durán, H.
Filiación:Departamento de Micro y Nanotecnología, Comisión Nacional de Energía Atómica, Av. General Paz 1499, (B1650KNA) San Martín, Buenos Aires, Argentina
Departamento de Radiobiología, Comisión Nacional de Energía Atómica, San Martín, Buenos Aires, Argentina
Gerencia de Investigación y Aplicaciones, Comisión Nacional de Energía Atómica, San Martín, Buenos Aires, Argentina
Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina
Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Escuela de Ciencia y Tecnología, Universidad Nacional de San Martín, San Martín, Buenos Aires, Argentina
Palabras clave:Catalytic subunits; Chinese Hamster ovary cells; DNA double-strand breaks; DNA-dependent protein kinase; Double-strand breaks; High linear energy transfers; Methods and materials; Nonhomologous end joining; Cell culture; DNA; Enzymes; Irradiation; Phosphorylation; Proteins; Repair; Radiation; deoxyribonucleic acid protein kinase; histone H2AX; lithium; protein kinase; proton; unclassified drug; animal cell; article; cell mutant; cell nucleus; cell survival; CHO cell; clonogenic assay; controlled study; DNA damage; DNA end joining repair; double stranded DNA break; enzyme active site; enzyme activity; female; gamma radiation; immunofluorescence; ionizing radiation; linear energy transfer; nonhuman; priority journal; protein dephosphorylation; protein phosphorylation; radiation dose; radiation response; radiosensitivity; Animals; Biological Markers; CHO Cells; Cricetinae; Cricetulus; DNA Breaks, Double-Stranded; DNA Repair; DNA-Activated Protein Kinase; Gamma Rays; Histones; Linear Energy Transfer; Radiation Dosage; Radiation Tolerance; Time Factors
Año:2013
Volumen:87
Número:4
Página de inicio:785
Página de fin:794
DOI: http://dx.doi.org/10.1016/j.ijrobp.2013.07.014
Título revista:International Journal of Radiation Oncology Biology Physics
Título revista abreviado:Int. J. Radiat. Oncol. Biol. Phys.
ISSN:03603016
CODEN:IOBPD
CAS:lithium, 7439-93-2; protein kinase, 9026-43-1; proton, 12408-02-5, 12586-59-3; Biological Markers; DNA-Activated Protein Kinase, 2.7.11.1; H2AFX protein, human; Histones
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03603016_v87_n4_p785_Bracalente

Referencias:

  • Rich, T., Allen, R.L., Wyllie, A.H., Defying death after DNA damage (2000) Nature, 407, pp. 777-783
  • Shrivastav, M., De Haro, L.P., Nickoloff, J.A., Regulation of DNA double-strand break repair pathway choice (2008) Cell Res, 18, pp. 134-147
  • Kurimasa, A., Kumano, S., Boubnov, N.V., Requirement for the kinase activity of human DNA-dependent protein kinase catalytic subunit in DNA strand break rejoining (1999) Mol Cell Biol, 19, pp. 3877-3884
  • Ding, Q., Reddy, Y.V., Wang, W., Autophosphorylation of the catalytic subunit of the DNA-dependent protein kinase is required for efficient end processing during DNA double-strand break repair (2003) Mol Cell Biol, 23, pp. 5836-5848
  • Hsu, F.M., Zhang, S., Chen, B.P., Role of DNA-dependent protein kinase catalytic subunit in cancer development and treatment (2012) Transl Cancer Res, 1, pp. 22-34
  • Redon, C.E., Nakamura, A.J., Zhang, Y.W., Histone gamma H2AX and poly(ADP-ribose) as clinical pharmacodynamic biomarkers (2010) Clin Cancer Res, 16, pp. 4532-4542
  • Rogakou, E.P., Pilch, D.R., Orr, A.H., DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139 (1998) J Biol Chem, 273, pp. 5858-5868
  • Ibanez, I.L., Bracalente, C., Molinari, B.L., Induction and rejoining of DNA double strand breaks assessed by H2AX phosphorylation in melanoma cells irradiated with proton and lithium beams (2009) Int J Radiat Oncol Biol Phys, 74, pp. 1226-1235
  • Rothkamm, K., Lobrich, M., Evidence for a lack of DNA double-strand break repair in human cells exposed to very low x-ray doses (2003) Proc Natl Acad Sci U S A, 100, pp. 5057-5062
  • Costes, S.V., Boissiere, A., Ravani, S., Imaging features that discriminate between foci induced by high- and low-LET radiation in human fibroblasts (2006) Radiat Res, 165, pp. 505-515
  • Orecchia, R., Zurlo, A., Loasses, A., Particle beam therapy (hadrontherapy): Basis for interest and clinical experience (1998) Eur J Cancer, 34, pp. 459-468
  • Priestley, A., Beamish, H.J., Gell, D., Molecular and biochemical characterisation of DNA-dependent protein kinase-defective rodent mutant irs-20 (1998) Nucleic Acids Res, 26, pp. 1965-1973
  • Belli, M., Bettega, D., Calzolari, P., Inactivation of human normal and tumour cells irradiated with low energy protons (2000) Int J Radiat Biol, 76, pp. 831-839
  • Tobias, C.A., Blakely, E.A., Chang, P.Y., Response of sensitive human ataxia and resistant t-1 cell lines to accelerated heavy ions (1984) Br J Cancer Suppl, 6, pp. 175-185
  • Kinashi, Y., Takahashi, S., Kashino, G., DNA double-strand break induction in ku80-deficient CHO cells following boron neutron capture reaction (2011) Radiat Oncol, 6, p. 106
  • Vandersickel, V., Depuydt, J., Van Bockstaele, B., Early increase of radiation-induced gamma H2AX foci in a human ku70/80 knockdown cell line characterized by an enhanced radiosensitivity (2010) J Radiat Res, 51, pp. 633-641
  • Mladenov, E., Iliakis, G., Induction and repair of DNA double strand breaks: The increasing spectrum of non-homologous end joining pathways (2011) Mutat Res, 711, pp. 61-72
  • Bourton, E.C., Plowman, P.N., Zahir, S.A., Multispectral imaging flow cytometry reveals distinct frequencies of gamma-H2AX foci induction in DNA double strand break repair defective human cell lines (2012) Cytometry A, 81, pp. 130-137
  • Peddi, P., Loftin, C.W., Dickey, J.S., DNA-PKCs deficiency leads to persistence of oxidatively induced DNA lesions in human tumor cells (2010) Free Radic Biol Med, 48, pp. 1435-1443
  • Anderson, J.A., Harper, J.V., Cucinotta, F.A., Participation of DNA-PKcs in DSB repair after exposure to high- and low-LET radiation (2010) Radiat Res, 174, pp. 195-205
  • Asaithamby, A., Hu, B., Chen, D.J., Unrepaired clustered DNA lesions induce chromosome breakage in human cells (2011) Proc Natl Acad Sci U S A, 108, pp. 8293-8298

Citas:

---------- APA ----------
Bracalente, C., Ibañez, I.L., Molinari, B., Palmieri, M., Kreiner, A., Valda, A., Davidson, J.,..., Durán, H. (2013) . Induction and persistence of large γh2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells. International Journal of Radiation Oncology Biology Physics, 87(4), 785-794.
http://dx.doi.org/10.1016/j.ijrobp.2013.07.014
---------- CHICAGO ----------
Bracalente, C., Ibañez, I.L., Molinari, B., Palmieri, M., Kreiner, A., Valda, A., et al. "Induction and persistence of large γh2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells" . International Journal of Radiation Oncology Biology Physics 87, no. 4 (2013) : 785-794.
http://dx.doi.org/10.1016/j.ijrobp.2013.07.014
---------- MLA ----------
Bracalente, C., Ibañez, I.L., Molinari, B., Palmieri, M., Kreiner, A., Valda, A., et al. "Induction and persistence of large γh2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells" . International Journal of Radiation Oncology Biology Physics, vol. 87, no. 4, 2013, pp. 785-794.
http://dx.doi.org/10.1016/j.ijrobp.2013.07.014
---------- VANCOUVER ----------
Bracalente, C., Ibañez, I.L., Molinari, B., Palmieri, M., Kreiner, A., Valda, A., et al. Induction and persistence of large γh2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells. Int. J. Radiat. Oncol. Biol. Phys. 2013;87(4):785-794.
http://dx.doi.org/10.1016/j.ijrobp.2013.07.014