Radiation induced DNA damage and damage repair in human tumor and fibroblast cell lines assessed by histone H2AX phosphorylation.
OBJECTIVE
To analyze the radiation-induced levels of gammaH2AX and its decay kinetics in 10 human cell lines covering a wide range of cellular radiosensitivity (SF2, 0.06-0.63).


METHODS
Five tumor cell lines included Colo-800 melanoma, two glioblastoma (MO59J and MO59K), fibrosarcoma HT 1080, and breast carcinoma MCF7. Five primary skin fibroblasts lines included two normal strains, an ataxia telangiectasia strain, and two fibroblast strains from breast cancer patients with an adverse early skin reaction to radiotherapy. Cellular radiosensitivity was assessed by colony-forming test. Deoxyribonucleic acid damage and repair were analyzed according to nuclear gammaH2AX foci intensity, with digital image analysis.


RESULTS
The cell lines tested showed a wide degree of variation in the background intensity of immunostained nuclear histone gammaH2AX, which was higher for the tumor cell lines compared with the fibroblast strains. It was not possible to predict clonogenic cell survival (SF2) for the 10 cell lines studied from the radiation-induced gammaH2AX intensity. In addition, the slopes of the dose-response (0-4 Gy) curves, the rates of gammaH2AX disappearance, and its residual expression (<or=18 h after irradiation) did not correlate with SF2 values.


CONCLUSIONS
The results from 10 cell lines showed that measurements of immunofluorescence intensity by digital image analysis of phosphorylated histone H2AX as a surrogate marker of DNA double-strand breaks did not allow reliable ranking of cell strains according to their clonogenic survival after irradiation.
Mahrhofer
Hartmut
H
Klinik für Strahlentherapie der Universität Würzburg, Würzburg, Germany.
Bürger
Susann
S
Oppitz
Ulrich
U
Flentje
Michael
M
Djuzenova
Cholpon S
CS
D002460:Cell Line / Q000528:radiation effects; D045744:Cell Line, Tumor / Q000528:radiation effects; D002467:Cell Nucleus / Q000378:metabolism; D003114:Colony-Forming Units Assay; D004249:DNA Damage; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000528:radiation effects; D006657:Histones / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D011836:Radiation Tolerance
C468783:H2AX protein, human; D006657:Histones