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Radiosensitisation of bladder cancer cells by panobinostat is modulated by Ku80 expression.

Groselj B, Kerr M, Kiltie AE - Radiother Oncol (2013)

Bottom Line: Resolution of γH2AX foci was determined by immunofluorescence confocal microscopy, cell cycle progression by FACS analysis and protein expression by western blotting.PAN had a greater radiosensitising effect in Ku80KD than RT112 or RAD51KD cells; enhancement ratios 1.35 for Ku80KD at 10nM (IC(20) for Ku80KD) and 1.31 for RT112 and RAD51KD at 25 nM (IC(40) for both).As muscle-invasive bladder tumours have reduced Ku-DNA binding, PAN could be particularly useful as a radiosensitiser in bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Gray Institute for Radiation Oncology and Biology, Department of Oncology, University of Oxford, United Kingdom.

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(a) Clonogenic assays in RT112, RAD51KD and Ku80KD cells treated with increasing ionising radiation doses and 10 nM or 25 nM PAN; (b) Percentages of cells surviving after 6 Gy, quantified from (a); (c) quantification of γH2AX foci from (d); (d) Confocal images of γH2AX foci (top panels) and DAPI stained cells (bottom panels) for cells treated with and without 5 Gy IR and 25 nM PAN.
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f0010: (a) Clonogenic assays in RT112, RAD51KD and Ku80KD cells treated with increasing ionising radiation doses and 10 nM or 25 nM PAN; (b) Percentages of cells surviving after 6 Gy, quantified from (a); (c) quantification of γH2AX foci from (d); (d) Confocal images of γH2AX foci (top panels) and DAPI stained cells (bottom panels) for cells treated with and without 5 Gy IR and 25 nM PAN.

Mentions: Cells were treated at 10 nM and 25 nM PAN concentrations, approximately equivalent to RT112’s IC10 and IC40. PAN significantly sensitised Ku80KD cells at 10 nM (SER = 1.35, P = 0.04) with less marked effects in parental RT112 cells (SER = 1.16, P = 0.05) and RAD51KD cells (SER = 1.22, P = 0.04). At 25 nM PAN (IC40), the SER for both RT112 and RAD51KD cells was 1.31 (P = 0.01 and P = 0.03, respectively), but this was higher for Ku80KD cells at a Ku80KD IC20 equivalent dose (1.56, P = 0.05; Fig. 2A and B).


Radiosensitisation of bladder cancer cells by panobinostat is modulated by Ku80 expression.

Groselj B, Kerr M, Kiltie AE - Radiother Oncol (2013)

(a) Clonogenic assays in RT112, RAD51KD and Ku80KD cells treated with increasing ionising radiation doses and 10 nM or 25 nM PAN; (b) Percentages of cells surviving after 6 Gy, quantified from (a); (c) quantification of γH2AX foci from (d); (d) Confocal images of γH2AX foci (top panels) and DAPI stained cells (bottom panels) for cells treated with and without 5 Gy IR and 25 nM PAN.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3824066&req=5

f0010: (a) Clonogenic assays in RT112, RAD51KD and Ku80KD cells treated with increasing ionising radiation doses and 10 nM or 25 nM PAN; (b) Percentages of cells surviving after 6 Gy, quantified from (a); (c) quantification of γH2AX foci from (d); (d) Confocal images of γH2AX foci (top panels) and DAPI stained cells (bottom panels) for cells treated with and without 5 Gy IR and 25 nM PAN.
Mentions: Cells were treated at 10 nM and 25 nM PAN concentrations, approximately equivalent to RT112’s IC10 and IC40. PAN significantly sensitised Ku80KD cells at 10 nM (SER = 1.35, P = 0.04) with less marked effects in parental RT112 cells (SER = 1.16, P = 0.05) and RAD51KD cells (SER = 1.22, P = 0.04). At 25 nM PAN (IC40), the SER for both RT112 and RAD51KD cells was 1.31 (P = 0.01 and P = 0.03, respectively), but this was higher for Ku80KD cells at a Ku80KD IC20 equivalent dose (1.56, P = 0.05; Fig. 2A and B).

Bottom Line: Resolution of γH2AX foci was determined by immunofluorescence confocal microscopy, cell cycle progression by FACS analysis and protein expression by western blotting.PAN had a greater radiosensitising effect in Ku80KD than RT112 or RAD51KD cells; enhancement ratios 1.35 for Ku80KD at 10nM (IC(20) for Ku80KD) and 1.31 for RT112 and RAD51KD at 25 nM (IC(40) for both).As muscle-invasive bladder tumours have reduced Ku-DNA binding, PAN could be particularly useful as a radiosensitiser in bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Gray Institute for Radiation Oncology and Biology, Department of Oncology, University of Oxford, United Kingdom.

Show MeSH
Related in: MedlinePlus