Normal expression of DNA repair proteins, hMre11, Rad50 and Rad51 but protracted formation of Rad50 containing foci in X-irradiated skin fibroblasts from radiosensitive cancer patients.
About 5% of oncology patients treated by radiation therapy develop acute or late radiotoxic effects whose molecular mechanisms remain poorly understood. In this study, we evaluated the potential role of DNA repair proteins in the hypersensitivity of cancer patients to radiation therapy. The expression levels and focal nuclear distribution of DNA repair proteins, hMre11, Rad50 and Rad51 were investigated in skin fibroblasts strains derived from cancer patients with adverse early skin reaction to radiotherapy using Western blot and foci immunofluorescence techniques, respectively. Cells from cancer patients with normal reaction to radiotherapy as well as cells from apparently healthy subjects served as controls. Cellular radiosensitivity after in vitro irradiation was assessed by the clonogenic survival assay. The clonogenic survival assay and Western blot analysis of the DNA repair proteins did not reveal any abnormalities in cellular radiosensitivity in vitro and in protein expression levels or their migration patterns in the fibroblasts derived from cancer patients with hypersensitive reaction to radiotherapy. In contrast, in vitro irradiated cells from radiosensitive patients exhibited a significantly higher number of nuclei with focally concentrated Rad50 protein than in both control groups. The observed alteration of the distribution of radiation-induced Rad50 foci in cells derived from cancer patients with acute side reactions to radiotherapy might contribute to their radiation therapy outcome. These data suggest the usefulness of the Rad50 foci analysis for predicting clinical response of cancer patients to radiotherapy.
Djuzenova
C
C
Klinik für Strahlentherapie der Universität Würzburg, Josef-Schneider-Strasse 11, D-97080 Würzburg, Germany. djuzenova_t@klinik.uni-wuerzburg.de
Mühl
B
B
Schakowski
R
R
Oppitz
U
U
Flentje
M
M
D017766:Acid Anhydride Hydrolases; D001706:Biopsy; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000532:radiotherapy; D018929:Cell Culture Techniques; D004260:DNA Repair; D045643:DNA Repair Enzymes / Q000096:biosynthesis*; D004268:DNA-Binding Proteins / Q000096:biosynthesis*; D005260:Female; D005347:Fibroblasts / Q000502:physiology; D020869:Gene Expression Profiling; D006801:Humans; D000076228:MRE11 Homologue Protein; D051135:Rad51 Recombinase; D011832:Radiation Injuries / Q000235:genetics* / Q000503:physiopathology*; D011836:Radiation Tolerance / Q000235:genetics*; D012867:Skin / Q000473:pathology
D004268:DNA-Binding Proteins; C121766:MRE11 protein, human; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D000076228:MRE11 Homologue Protein; D017766:Acid Anhydride Hydrolases; C112158:RAD50 protein, human; D045643:DNA Repair Enzymes