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Lentivirus-mediated RNA interference of Ku70 to enhance radiosensitivity of human mammary epithelial cells

Veerle Vandersickel UGent, M Mancini, E Marras, P Willems, J Slabbert, Jan Philippé UGent, Ellen Deschepper UGent, Hubert Thierens UGent, G Perletti and Anne Vral UGent (2010) INTERNATIONAL JOURNAL OF RADIATION BIOLOGY. 86(2). p.114-124
abstract
Purpose: To investigate the radiosensitising effect of Ku autoantigen 70 (Ku70) and Ku autoantigen 80 (Ku80) knockdown by lentivirus-mediated RNA interference (RNAi) in the MCF10A immortalised human mammary epithelial cell line. Materials and methods: MCF10A cells were infected with lentiviral vector, for RNAi of Ku70. The Ku70-knockdown cell line (Ku70i) and a mock-infected control cell line (LVTHM) were used to perform radiation experiments. For the in vitro Micronucleus (MN) assay, both cell lines were irradiated with doses of 2 and 4 Gy (CO)-C-60 gamma-rays. For cell survival experiments, doses ranging between 0 and 8 Gy were used. Results: Western blot analysis showed that the Ku70 lentiviral vector was effective in silencing the expression of both Ku70 and Ku80. A significantly higher radiation-induced MN yield was obtained in the Ku70i cell line compared to the control LVTHM cell line. RNAi of Ku70 also resulted in a lower survival yield after irradiation compared to the control cell line. Analysis of cell death mechanisms showed that MCF10A cells (Ku70i and LVTHM) do not undergo apoptosis, but undergo Post-irradiation cellular senescence. Conclusion: RNTAi of Ku70 resulted in increased chromosomal and cellular radiosensitivity in the MCF10A human mammary cell line after irradiation with (CO)-C-60 gamma-rays. These results further strengthen the role of the Ku protein in correct DNA double strand break (DSB) repair.
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author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
CHROMOSOMAL RADIOSENSITIVITY, ACCELERATED SENESCENCE, IN-VITRO, END-JOINING GENES, DOMINANT-NEGATIVE KU70, BREAST-CANCER PATIENTS, GAMMA-RADIATION, IONIZING-RADIATION, HUMAN FIBROBLASTS, TUMOR-CELLS
journal title
INTERNATIONAL JOURNAL OF RADIATION BIOLOGY
Int. J. Radiat. Biol.
volume
86
issue
2
pages
11 pages
publisher
TAYLOR & FRANCIS LTD
place of publication
ABINGDON
Web of Science type
Article
Web of Science id
000274629400004
JCR category
NUCLEAR SCIENCE & TECHNOLOGY
JCR impact factor
1.861 (2010)
JCR rank
2/34 (2010)
JCR quartile
1 (2010)
ISSN
0955-3002
DOI
10.3109/09553000903419940
language
English
UGent publication?
yes
classification
A1
copyright statement
I don't know the status of the copyright for this publication
id
920715
handle
http://hdl.handle.net/1854/LU-920715
date created
2010-04-02 14:49:20
date last changed
2010-04-08 13:30:11
@article{920715,
  abstract     = {Purpose: To investigate the radiosensitising effect of Ku autoantigen 70 (Ku70) and Ku autoantigen 80 (Ku80) knockdown by lentivirus-mediated RNA interference (RNAi) in the MCF10A immortalised human mammary epithelial cell line.
Materials and methods: MCF10A cells were infected with lentiviral vector, for RNAi of Ku70. The Ku70-knockdown cell line (Ku70i) and a mock-infected control cell line (LVTHM) were used to perform radiation experiments. For the in vitro Micronucleus (MN) assay, both cell lines were irradiated with doses of 2 and 4 Gy (CO)-C-60 gamma-rays. For cell survival experiments, doses ranging between 0 and 8 Gy were used.

Results: Western blot analysis showed that the Ku70 lentiviral vector was effective in silencing the expression of both Ku70 and Ku80. A significantly higher radiation-induced MN yield was obtained in the Ku70i cell line compared to the control LVTHM cell line. RNAi of Ku70 also resulted in a lower survival yield after irradiation compared to the control cell line. Analysis of cell death mechanisms showed that MCF10A cells (Ku70i and LVTHM) do not undergo apoptosis, but undergo Post-irradiation cellular senescence.

Conclusion: RNTAi of Ku70 resulted in increased chromosomal and cellular radiosensitivity in the MCF10A human mammary cell line after irradiation with (CO)-C-60 gamma-rays. These results further strengthen the role of the Ku protein in correct DNA double strand break (DSB) repair.},
  author       = {Vandersickel, Veerle and Mancini, M and Marras, E and Willems, P and Slabbert, J and Philipp{\'e}, Jan and Deschepper, Ellen and Thierens, Hubert and Perletti, G and Vral, Anne},
  issn         = {0955-3002},
  journal      = {INTERNATIONAL JOURNAL OF RADIATION BIOLOGY},
  keyword      = {CHROMOSOMAL RADIOSENSITIVITY,ACCELERATED SENESCENCE,IN-VITRO,END-JOINING GENES,DOMINANT-NEGATIVE KU70,BREAST-CANCER PATIENTS,GAMMA-RADIATION,IONIZING-RADIATION,HUMAN FIBROBLASTS,TUMOR-CELLS},
  language     = {eng},
  number       = {2},
  pages        = {114--124},
  publisher    = {TAYLOR \& FRANCIS LTD},
  title        = {Lentivirus-mediated RNA interference of Ku70 to enhance radiosensitivity of human mammary epithelial cells},
  url          = {http://dx.doi.org/10.3109/09553000903419940},
  volume       = {86},
  year         = {2010},
}

Chicago
Vandersickel, Veerle, M Mancini, E Marras, P Willems, J Slabbert, Jan Philippé, Ellen Deschepper, Hubert Thierens, G Perletti, and Anne Vral. 2010. “Lentivirus-mediated RNA Interference of Ku70 to Enhance Radiosensitivity of Human Mammary Epithelial Cells.” International Journal of Radiation Biology 86 (2): 114–124.
APA
Vandersickel, V., Mancini, M., Marras, E., Willems, P., Slabbert, J., Philippé, J., Deschepper, E., et al. (2010). Lentivirus-mediated RNA interference of Ku70 to enhance radiosensitivity of human mammary epithelial cells. INTERNATIONAL JOURNAL OF RADIATION BIOLOGY, 86(2), 114–124.
Vancouver
1.
Vandersickel V, Mancini M, Marras E, Willems P, Slabbert J, Philippé J, et al. Lentivirus-mediated RNA interference of Ku70 to enhance radiosensitivity of human mammary epithelial cells. INTERNATIONAL JOURNAL OF RADIATION BIOLOGY. ABINGDON: TAYLOR & FRANCIS LTD; 2010;86(2):114–24.
MLA
Vandersickel, Veerle, M Mancini, E Marras, et al. “Lentivirus-mediated RNA Interference of Ku70 to Enhance Radiosensitivity of Human Mammary Epithelial Cells.” INTERNATIONAL JOURNAL OF RADIATION BIOLOGY 86.2 (2010): 114–124. Print.