Background
Little is known about the cellular effects of exposure to mixed beams of high and low linear energy transfer radiation. So far, the effects of combined exposures have mainly been assessed with clonogenic survival or cytogenetic methods, and the results are contradictory. The gamma-H2AX assay has up to now not been applied in this context, and it is a promising tool for investigating the early cellular response to mixed beam irradiation.

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Gamma-H2AX foci in cells exposed to a mixed beam of X-rays and alpha particles

The purpose of this study was to analyse the cytogenetic effect of exposing human peripheral blood lymphocytes (PBL) to a mixed beam of alpha particles and X-rays. Whole blood collected from one donor was exposed to different doses of alpha particles ((241)Am), X-rays and a combination of both. All exposures were carried out at 37 °C. Three independent experiments were performed. Micronuclei (MN) in binucleated PBL were scored as the endpoint. Moreover, the size of MN was measured. The results show that exposure of PBL to a mixed beam of high and low linear energy transfer radiation led to significantly higher than expected frequencies of MN. The measurement of MN size did not reveal any differences between the effect of alpha particles and mixed beam. In conclusion, a combined exposure of PBL to alpha particles and X-rays leads to a synergistic effect as measured by the frequency of MN. From the analysis of MN distributions, we conclude that the increase was due to an impaired repair of X-ray-induced DNA damage.

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Cellular effects after exposure to mixed beams of ionizing radiation

Predicting the risk of second malignant neoplasms is complicated by uncertainties regarding the shape of the dose–response relationship at high doses. Limited understanding of the competitive relationship between cell killing and the accumulation of DNA lesions at high doses, as well as the effects of other modulatory factors unique to radiation exposure during radiotherapy, such as dose heterogeneity across normal tissue and dose fractionation, contribute to these uncertainties. The aim of this study was to analyze the impact of fractionated irradiations on two cell systems, focusing on the endpoints relevant for cancer induction. To simulate the heterogeneous dose distribution across normal tissue during radiotherapy, exponentially growing VH10 fibroblasts and AHH-1 lymphoblasts were irradiated with 9 and 12 fractions (VH10) and 10 fractions (AHH-1) at 0.25, 0.5, 1, or 2 Gy per fraction. The effects on cell growth, cell survival, radiosensitivity and the accumulation of residual DNA damage lesions were analyzed as functions of dose per fraction and the total absorbed dose. Residual γH2AX foci and other DNA damage markers (micronuclei, nuclear buds, and giant nuclei) were accumulated at high doses in both cell types, but in a cell type-dependent manner. The competitive relationship between cell killing and the accumulation of carcinogenic DNA damage following multifractional radiation exposure is cell type-specific.

/pdf/cell-type-specific-patterns-in-the-accumulation-of-dna-1gbcw5od.pdf

Cell Type-Specific Patterns in the Accumulation of DNA Damage Following Multifractional Radiation Exposure

The aim of this study was to evaluate the DNA damage induced by low and high linear energy transfer radiation in two related cell lines with different radiation sensitivity, CHO10B2 and irs-20 cells (defective in DNA-PKcs). Double-strand breaks were assessed by the detection of phosphorylated histone H2AX foci (γH2AX). The number of foci increased as a function of dose for both cell lines and types of radiation. However, irs-20 cells showed a higher number of foci than the parental CHO10B2. An increase in the foci size was observed for both cell lines after lithium irradiation. This could be attributed to clusters of DNA damage. Moreover, the number of lithium-induced larger foci per nuclei increased with dose and fitted with the expected number of hits per nucleus. Concluding, γH2AX foci size provides a potential tool to characterise the intrinsic radiosensitivity of cells related to DNA damage and to compare the effects of different quality radiations.

Assessment of γH2AX nuclear foci number and size in normal and repair-deficient cells irradiated with low and high linear energy transfer radiation

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Assessment of γH2AX nuclear foci number and size in normal and repair-deficient cells irradiated with low and high linear energy transfer radiation