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

PURPOSE

To assess the potential of using the residual phosphorylation of histone H2AX (gammaH2AX) after irradiation as a marker of radiosensitivity in vitro.

MATERIAL AND METHODS

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.

RESULTS

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 <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.

CONCLUSION

In the two cell lines investigated the mean number of residual foci of gammaH2AX can be used to predict differences in the radiation dose response relationship in vitro.