Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology · 2011
DNA methylation changes in cells regrowing after fractioned ionizing radiation.
Сравнительное исследованиеСтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
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Kuhmann Christine, Weichenhan Dieter, Rehli Michael, Plass Christoph, Schmezer Peter, Popanda Odilia
Аннотация
BACKGROUND AND PURPOSE: Repeated exposure to ionizing radiation (IR) can result in adaptive reactions. While DNA methylation changes in adaption to repeated stress exposure are established for a variety of drugs, their role in fractioned ionizing radiation is largely unknown.
MATERIAL AND METHODS: MCF7 breast cancer cells were treated 5 times a week with IR in fractions of 2 Gy, resulting in total doses of 10 and 20 Gy. Cells were harvested 48 and 72 h after the last irradiation, as well as after a recovery period of at least 14 d. To identify genes differentially methylated in irradiated versus non-irradiated cells, we used methyl-CpG immunoprecipitation (MCIp) followed by global methylation profiling on CpG island microarrays.
RESULTS: MCIp profiling revealed methylation changes in several CpG islands 48 h after FIR with 10 and 20 Gy. Cells receiving a total dose of 10 Gy started regrowing after 14 d and exhibited similar radioresistance as mock-treated cells. Differential methylation of the CpG units associated with FOXC1 (p<0.001) and TRAPPC9 (p<0.001) could be confirmed by time-of-flight mass spectrometry (Sequenom).
CONCLUSIONS: In summary, these data indicate that regrowth of MCF7 cells after 10 Gy FIR is associated with locus-specific alterations in DNA methylation.
MATERIAL AND METHODS: MCF7 breast cancer cells were treated 5 times a week with IR in fractions of 2 Gy, resulting in total doses of 10 and 20 Gy. Cells were harvested 48 and 72 h after the last irradiation, as well as after a recovery period of at least 14 d. To identify genes differentially methylated in irradiated versus non-irradiated cells, we used methyl-CpG immunoprecipitation (MCIp) followed by global methylation profiling on CpG island microarrays.
RESULTS: MCIp profiling revealed methylation changes in several CpG islands 48 h after FIR with 10 and 20 Gy. Cells receiving a total dose of 10 Gy started regrowing after 14 d and exhibited similar radioresistance as mock-treated cells. Differential methylation of the CpG units associated with FOXC1 (p<0.001) and TRAPPC9 (p<0.001) could be confirmed by time-of-flight mass spectrometry (Sequenom).
CONCLUSIONS: In summary, these data indicate that regrowth of MCF7 cells after 10 Gy FIR is associated with locus-specific alterations in DNA methylation.
Предметные рубрики MeSH
Adaptation, PhysiologicalBreast NeoplasmsCell ProliferationCpG IslandsDNA MethylationDose Fractionation, RadiationDose-Response Relationship, RadiationFemaleGene Expression ProfilingHumansImmunoprecipitationMicroarray AnalysisRadiation, IonizingReference ValuesSensitivity and SpecificityTumor Cells, Cultured