X-ray-induced DNA double-strand breaks in mouse l1210 cells: a new computational method for analyzing neutral filter elution data.
The aim of this article is to present a method for studying the shape of the dose and repair responses for X-ray-induced double-strand breaks (DSBs) as measured by neutral filter elution (NFE). The approach is closely related to a method we developed for the use of specific molecular size markers and used for determination of the absolute number of randomly distributed radiation-induced DSBs by pulsed-field gel electrophoresis (PFGE). Mouse leukemia L1210 cells were X-irradiated with 0-50 Gy. Samples were then evaluated both with PFGE and with NFE. Assuming that with both migration (PFGE) and elution (NFE), a heterogeneous population of double-stranded DNA fragments will start with the smallest fragments and proceed with increasingly larger fragments, it is possible to match the migration behavior of fractions of fragments smaller than a certain size to the fraction eluted at a specific time. This assumption does not exclude the possibility of DNA being sheared in the NFE filter. The yield, as determined by the size markers in PFGE, was used to find the corresponding elution times in the NFE experiment. These experimentally used elution times could then reversely be interpreted as size markers which finally were used to calculate DSBs/Mbp as a function of X-ray dose. The resulting lines were almost straight. The data were also plotted as relative elution and showed that, as expected, the dose response then appears with a more pronounced sigmoid shape.
Cedervall
Björn
B
Medical Radiation Biology, Department of Oncology and Pathology, Karolinska Institutet, Box 260, SE-171 76 Stockholm, Sweden. cedervall@raffys.ki.se
Edgren
Margareta R
MR
Lewensohn
Rolf
R
D000818:Animals; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000302:isolation & purification / Q000528:radiation effects*; D004307:Dose-Response Relationship, Radiation; D016521:Electrophoresis, Gel, Pulsed-Field; D005374:Filtration / Q000379:methods*; D007939:Leukemia L1210 / Q000473:pathology*; D008433:Mathematics; D051379:Mice; D008970:Molecular Weight; D011897:Random Allocation; D012212:Rheology; D013997:Time Factors; D014407:Tumor Cells, Cultured / Q000528:radiation effects
D004273:DNA, Neoplasm