Prediction of clonogenic cell survival curves based on the number of residual DNA double strand breaks measured by gammaH2AX staining

To assess the potential of using the residual phosphorylation of histone H2AX (gammaH2AX) after irradiation as a marker of radiosensitivity in vitro. Confluent cell cultures of FaDu and SKX human squamous cell carcinoma lines were irradiated with graded single doses. Twenty-four hours after irradiat...

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Published in:International journal of radiation biology 2009-11, Vol.85 (11), p.1032
Main Authors: Menegakis, Apostolos, Yaromina, Ala, Eicheler, Wolfgang, Dörfler, Annegret, Beuthien-Baumann, Bettina, Thames, Howard D, Baumann, Michael, Krause, Mechthild
Format: Article
Language:English
Subjects:
Carcinoma, Squamous Cell - metabolism
Carcinoma, Squamous Cell - pathology
Carcinoma, Squamous Cell - radiotherapy
Cell Line, Tumor
Cell Survival - radiation effects
DNA Breaks, Double-Stranded
DNA, Neoplasm - radiation effects
Dose-Response Relationship, Radiation
Histones - metabolism
Humans
Immunohistochemistry
Radiation Tolerance
Tumor Stem Cell Assay
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container_issue 11
container_start_page 1032
container_title International journal of radiation biology
container_volume 85
creator Menegakis, Apostolos
Yaromina, Ala
Eicheler, Wolfgang
Dörfler, Annegret
Beuthien-Baumann, Bettina
Thames, Howard D
Baumann, Michael
Krause, Mechthild
description To assess the potential of using the residual phosphorylation of histone H2AX (gammaH2AX) after irradiation as a marker of radiosensitivity in vitro. Confluent cell cultures of FaDu and SKX human squamous cell carcinoma lines were irradiated with graded single doses. Twenty-four hours after irradiation cells were seeded for standard colony forming assay (CFA). In parallel, staining for gammaH2AX was performed to visualise the residual foci. In the CFA, FaDu showed a higher radioresistance than SKX. After analysis of the residual foci data, we constructed 'predicted' survival curves using two different methods. First, the proportion of nuclei with
doi_str_mv 10.3109/09553000903242149
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Confluent cell cultures of FaDu and SKX human squamous cell carcinoma lines were irradiated with graded single doses. Twenty-four hours after irradiation cells were seeded for standard colony forming assay (CFA). In parallel, staining for gammaH2AX was performed to visualise the residual foci. In the CFA, FaDu showed a higher radioresistance than SKX. After analysis of the residual foci data, we constructed 'predicted' survival curves using two different methods. First, the proportion of nuclei with &lt;3 foci was found to correlate closely with the observed surviving fraction (SF) in FaDu, with a slight overestimation of the true SF in SKX. Second, there was a strong linear correlation of the mean number of residual foci and observed -lnSF. Based on regression analysis, we calculated the SF for both cell lines based on the mean number of residual gammaH2AX foci. This second approach again led to a good correlation of predicted and observed SF values in FaDu and a (slight) overestimation in SKX. 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subjects Carcinoma, Squamous Cell - metabolism
Carcinoma, Squamous Cell - pathology
Carcinoma, Squamous Cell - radiotherapy
Cell Line, Tumor
Cell Survival - radiation effects
DNA Breaks, Double-Stranded
DNA, Neoplasm - radiation effects
Dose-Response Relationship, Radiation
Histones - metabolism
Humans
Immunohistochemistry
Radiation Tolerance
Tumor Stem Cell Assay
title Prediction of clonogenic cell survival curves based on the number of residual DNA double strand breaks measured by gammaH2AX staining
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