Assessment of light scatter by nucleoids as a rapid predictive assay of radiosensitivity

Abstract

A study has been made of the practicality of using the assay of light scatter by nucleoids as a rapid predictive test of cellular radiosensitivity. With this technique the effect of irradiation on DNA organization is measured using flow cytometry after staining irradiated nucleoids with a high concentration of ethidium bromide. Damaged nucleoids fail to respond to the ethidium bromide-induced contraction and scatter more forward-angle light than less damaged nucleoids. Seventeen different cell lines were assessed using a single lysis condition and radiation dose. Significant differences in the levels of radiation-induced forward-angle light scatter by nucleoids were seen between CHO cells and cells of two radiosensitive mutant cell lines (xrs-6, EM9), and between cells of two ovarian carcinoma lines that showed marked differences in radiosensitivity measured using a clonogenic assay. However, other cell lines which differed in clonogenic radiosensitivity showed similar forward-angle light scatter by nucleoids. When all 17 cell lines were included in the analysis, there was no correlation between measurements of radiosensitivity by assays of clonogenicity and light scatter by nucleoids. In addition, although intraexperimental variation was small, the level of interexperimental variability was only slightly smaller (coefficient of variation of 13%) than the degree of heterogeneity observed between the different cell lines (coefficient of variation of 16%). These findings support the notion for a role of nuclear structure as a determinant of intrinsic radiosensitivity for some cell lines but suggest that for others there must be additional, more dominant factors.