Open Access Dissertation
Date of Award
James D. Tucker
The dose-effect relationships of cells exposed to ionizing radiation are frequently described by linear quadratic (LQ) models over an extended dose range. However, many mammalian cell lines, when acutely irradiated in G2 at doses below 30 cGy, show hyper-radiosensitivity (HRS) as measured by reduced clonogenic cell survival, thereby indicating greater cell lethality than is predicted by extrapolation from high-dose responses. We therefore hypothesized that the cytogenetic response in G2 cells to low doses would also be steeper than predicted by LQ extrapolation from high doses. We tested our hypothesis by exposing four normal human lymphoblastoid cell lines to 0 - 400 cGy of Cobalt-60 gamma radiation. The cytokinesis block micronucleus assay was used to determine the frequencies of micronuclei and nucleoplasmic bridges. To characterize the dependence of the cytogenetic damage on dose, univariate and multivariate regression analyses were used to compare the responses in the low- (HRS) and high-dose response regions. Our data indicate that the slope of the response for all four cell lines at doses below 20 cGy during G2 is greater than predicted by an LQ extrapolation from the high-dose responses for both micronuclei and bridges. Moreover, the cytogenetic effects of oxygen (O2) levels in tissue culture media on HRS have not been evaluated. We asked whether HRS was lost in G2-irradiated cells grown in atmospheres of 2.5% or 5% O2, compared to responses by cells cultured in ambient (~21%) O2. The results indicate a loss of HRS when cells are cultured and irradiated either in 2.5% or 5% O2. We then evaluated whether low O2 levels either before or after exposure were responsible for the loss of HRS. For cells irradiated in an atmosphere of 5% O2, subsequent immediate re-oxygenation to ambient O2 levels restored the HRS effect, while cells cultured and irradiated at ambient O2 levels and then transferred to 5% O2 exhibited little or no HRS, indicating that ambient O2 levels after, but not before, radiation substantially affect the amounts of cytogenetic damage. HRS was not observed when cells were irradiated in G1. At doses of 40 - 400 cGy there was significantly less cytogenetic damage when cells were recovering from radiation at low O2 levels than at ambient O2 levels. Here we provide the first cytogenetic evidence for the loss of HRS at low O2 levels in G2-irradiated cells; these results suggest that at low O2 levels for all doses evaluated there is either less damage to DNA, perhaps due to lower amounts of reactive oxygen species, or that DNA damage repair pathways are activated more efficiently.
Joshi, Gnanada Sameer, "Cytogenetic Evaluation Of Low Dose Hyper-Radiosensitivity In Human Lymphoblastoid Cell Lines Exposed To Cobalt-60 Gamma Radiation." (2014). Wayne State University Dissertations. 1074.