The potential value of the neutral comet assay and the expression of genes associated with DNA damage in assessing the radiosensitivity of tumor cells

► Potential of comet assay in assessing tumor radiosensitivity was evaluated. ► Neutral comet assay showed better correlation with radiosensitivity. ► It also correlated with radiosensitivity after fractionated dose of radiation. ► Expression of genes involved in DNA repair also correlated with radi...

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Published in:Mutation research. Genetic toxicology and environmental mutagenesis 2012-10, Vol.748 (1-2), p.52-59
Main Authors: Jayakumar, Sundarraj, Bhilwade, Hari N., Pandey, Badri N., Sandur, Santosh K., Chaubey, Ramesh C.
Format: Article
Language:English
Subjects:
Cells
Comet assay
DNA damage
Gene expression
Ionizing radiation
Mutation
Predictive assay
Radiation therapy
Radiosensitivity
Tumors
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Summary:► Potential of comet assay in assessing tumor radiosensitivity was evaluated. ► Neutral comet assay showed better correlation with radiosensitivity. ► It also correlated with radiosensitivity after fractionated dose of radiation. ► Expression of genes involved in DNA repair also correlated with radiosensitivity. The assessment of tumor radiosensitivity would be particularly useful in optimizing the radiation dose during radiotherapy. Therefore, the degree of correlation between radiation-induced DNA damage, as measured by the alkaline and the neutral comet assays, and the clonogenic survival of different human tumor cells was studied. Further, tumor radiosensitivity was compared with the expression of genes associated with the cellular response to radiation damage. Five different human tumor cell lines were chosen and the radiosensitivity of these cells was established by clonogenic assay. Alkaline and neutral comet assays were performed in γ-irradiated cells (2–8Gy; either acute or fractionated). Quantitative PCR was performed to evaluate the expression of DNA damage response genes in control and irradiated cells. The relative radiosensitivity of the cell lines assessed by the extent of DNA damage (neutral comet assay) immediately after irradiation (4Gy or 6Gy) was in agreement with radiosensitivity pattern obtained by the clonogenic assay. The survival fraction of irradiated cells showed a better correlation with the magnitude of DNA damage measured by the neutral comet assay (r=−0.9; P
ISSN:1383-5718
1879-3592
DOI:10.1016/j.mrgentox.2012.06.008