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Early increase of radiation-induced gamma H2AX foci in a human Ku70/80 knockdown cell line characterized by an enhanced radiosensitivity

Veerle Vandersickel UGent, Julie Depuydt UGent, Bram Van Bockstaele UGent, Gianpaolo Perletti, Jan Philippé UGent, Hubert Thierens UGent and Anne Vral UGent (2010) JOURNAL OF RADIATION RESEARCH. 51(6). p.633-641
abstract
A better understanding of the underlying mechanisms of DNA repair after exposure to ionizing radiation represents a research priority aimed at improving the outcome of clinical radiotherapy. Because of the close association with DNA double strand break (DSB) repair, phosphorylation of the histone H2AX protein (gamma H2AX), quantified by immunodetection, has recently been used as a method to study DSB induction and repair at low and clinically relevant radiation doses. However, the lack of consistency in literature points to the need to further validate the role of H2AX phosphorylation in DSB repair and the use of this technique to determine intrinsic radiosensitivity. In the present study we used human mammary epithelial MCF10A cells, characterized by a radiosensitive phenotype due to reduced levels of the Ku70 and Ku80 repair proteins, and investigated whether this repair-deficient cell line displays differences in the phosphorylation pattern of H2AX protein compared to repair-proficient MCF10A cells. This was established by measuring formation and disappearance of gamma H2AX foci after irradiating synchronized cell populations with Co-60 gamma-rays. Our results show statistically significant differences in the number of gamma H2AX foci between the repair-deficient and -proficient cell line, with a higher amount of gamma H2AX foci present at early times post-irradiation in the Ku-deficient cell line. However, the disappearance of those differences at later post-irradiation times questions the use of this assay to determine intrinsic radiosensitivity, especially in a clinical setting.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Ku70/80 heterodimer, DNA double strand breaks, Radiosensitivity, DOUBLE-STRAND BREAKS, HISTONE H2AX PHOSPHORYLATION, DNA-DAMAGE, IONIZING-RADIATION, TUMOR-CELLS, KU PROTEIN, IN-VIVO, X-RAYS, REPAIR, CHROMATIN, Ionizing radiation, H2AX phosphorylation
journal title
JOURNAL OF RADIATION RESEARCH
J. Radiat. Res.
volume
51
issue
6
pages
633 - 641
Web of Science type
Article
Web of Science id
000285174400002
JCR category
BIOLOGY
JCR impact factor
2.007 (2010)
JCR rank
29/84 (2010)
JCR quartile
2 (2010)
ISSN
0449-3060
DOI
10.1269/jrr.10033
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1146147
handle
http://hdl.handle.net/1854/LU-1146147
date created
2011-02-10 18:00:35
date last changed
2011-02-11 09:17:09
@article{1146147,
  abstract     = {A better understanding of the underlying mechanisms of DNA repair after exposure to ionizing radiation represents a research priority aimed at improving the outcome of clinical radiotherapy. Because of the close association with DNA double strand break (DSB) repair, phosphorylation of the histone H2AX protein (gamma H2AX), quantified by immunodetection, has recently been used as a method to study DSB induction and repair at low and clinically relevant radiation doses. However, the lack of consistency in literature points to the need to further validate the role of H2AX phosphorylation in DSB repair and the use of this technique to determine intrinsic radiosensitivity. In the present study we used human mammary epithelial MCF10A cells, characterized by a radiosensitive phenotype due to reduced levels of the Ku70 and Ku80 repair proteins, and investigated whether this repair-deficient cell line displays differences in the phosphorylation pattern of H2AX protein compared to repair-proficient MCF10A cells. This was established by measuring formation and disappearance of gamma H2AX foci after irradiating synchronized cell populations with Co-60 gamma-rays. Our results show statistically significant differences in the number of gamma H2AX foci between the repair-deficient and -proficient cell line, with a higher amount of gamma H2AX foci present at early times post-irradiation in the Ku-deficient cell line. However, the disappearance of those differences at later post-irradiation times questions the use of this assay to determine intrinsic radiosensitivity, especially in a clinical setting.},
  author       = {Vandersickel, Veerle and Depuydt, Julie and Van Bockstaele, Bram and Perletti, Gianpaolo and Philipp{\'e}, Jan and Thierens, Hubert and Vral, Anne},
  issn         = {0449-3060},
  journal      = {JOURNAL OF RADIATION RESEARCH},
  keyword      = {Ku70/80 heterodimer,DNA double strand breaks,Radiosensitivity,DOUBLE-STRAND BREAKS,HISTONE H2AX PHOSPHORYLATION,DNA-DAMAGE,IONIZING-RADIATION,TUMOR-CELLS,KU PROTEIN,IN-VIVO,X-RAYS,REPAIR,CHROMATIN,Ionizing radiation,H2AX phosphorylation},
  language     = {eng},
  number       = {6},
  pages        = {633--641},
  title        = {Early increase of radiation-induced gamma H2AX foci in a human Ku70/80 knockdown cell line characterized by an enhanced radiosensitivity},
  url          = {http://dx.doi.org/10.1269/jrr.10033},
  volume       = {51},
  year         = {2010},
}

Chicago
Vandersickel, Veerle, Julie Depuydt, Bram Van Bockstaele, Gianpaolo Perletti, Jan Philippé, Hubert Thierens, and Anne Vral. 2010. “Early Increase of Radiation-induced Gamma H2AX Foci in a Human Ku70/80 Knockdown Cell Line Characterized by an Enhanced Radiosensitivity.” Journal of Radiation Research 51 (6): 633–641.
APA
Vandersickel, V., Depuydt, J., Van Bockstaele, B., Perletti, G., Philippé, J., Thierens, H., & Vral, A. (2010). Early increase of radiation-induced gamma H2AX foci in a human Ku70/80 knockdown cell line characterized by an enhanced radiosensitivity. JOURNAL OF RADIATION RESEARCH, 51(6), 633–641.
Vancouver
1.
Vandersickel V, Depuydt J, Van Bockstaele B, Perletti G, Philippé J, Thierens H, et al. Early increase of radiation-induced gamma H2AX foci in a human Ku70/80 knockdown cell line characterized by an enhanced radiosensitivity. JOURNAL OF RADIATION RESEARCH. 2010;51(6):633–41.
MLA
Vandersickel, Veerle, Julie Depuydt, Bram Van Bockstaele, et al. “Early Increase of Radiation-induced Gamma H2AX Foci in a Human Ku70/80 Knockdown Cell Line Characterized by an Enhanced Radiosensitivity.” JOURNAL OF RADIATION RESEARCH 51.6 (2010): 633–641. Print.