Yeast DEL assay detects protection against radiation-induced cytotoxicity and genotoxicity: adaptation of a microtiter plate version.
The DEL assay in yeast detects DNA deletions that are inducible by many carcinogens. Here we use the colorimetric agent MTS to adapt the yeast DEL assay for microwell plate measurement of ionizing radiation-induced cell killing and DNA deletions. Using the microwell-based DEL assay, cell killing and genotoxic DNA deletions both increased with radiation dose between 0 and 2000 Gy. We used the microwell-based DEL assay to assess the effectiveness of varying concentrations of five different radioprotectors, N-acetyl-l-cysteine, l-ascorbic acid, DMSO, Tempol and Amifostine, and one radiosensitizer, 5-bromo-2-deoxyuridine. The microwell format of the DEL assay was able to successfully detect protection against and sensitization to both radiation-induced cytotoxicity and genotoxicity. Such radioprotection and sensitization detected by the microwell-based DEL assay was validated and compared with similar measurements made using the traditional agar-based assay format. The yeast DEL assay in microwell format is an effective tool for rapidly detecting chemical protectors and sensitizers to ionizing radiation and is automatable for chemical high-throughput screening purposes.
Department of Radiation Oncology, David Geffen School of Medicine at UCLA and UCLA School of Public Health, Los Angeles, California 90095, USA.
Kurt
K
Hafer
Yelena
Y
Rivina
Robert H
RH
Schiestl
D002470:Cell Survival / Q000528:radiation effects; D017353:Gene Deletion; D008624:Mercaptoethylamines / Q000494:pharmacology; D011837:Radiation-Protective Agents / Q000494:pharmacology*; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000528:radiation effects*
D008624:Mercaptoethylamines; D011837:Radiation-Protective Agents; D011838:Radiation-Sensitizing Agents; D017382:Reactive Oxygen Species; C020174:N-(2-mercaptoethyl)-1,3-diaminopropane