TY - JOUR
T1 - Melanoma cells express elevated levels of phosphorylated histone H2AX foci
AU - Warters, Raymond L.
AU - Adamson, Patrick J.
AU - Pond, Christopher D.
AU - Leachman, Sancy A.
N1 - Funding Information:
The authors thank the Cell Culture Core facility of Yale Skin Diseases Research Center (supported by USPH grant AR41942, R.E. Tigelaar Program Investigator) for providing the melanoma cell lines used in this report. We would also like to thank the Huntsman Cancer Institute for the use of the fluorescence microscope and the irradiation facility. We thank Dr Wayne Green, Director of the Health Sciences Center Flow Cytometry Facility, for his help in analysis of flow cytometry results. This research was supported by the Office of Science (BER), US Department of Energy, Grant number DE-FG03-01ER63240.
PY - 2005/4
Y1 - 2005/4
N2 - When human cells sustain a DNA double-strand break (dsb), histone H2AX in chromatin surrounding the DNA break is phosphorylated, marking repair foci. The number of phosphorylated histone H2AX (γH2AX) foci approximates the number of dsb present in the cell's nuclear DNA. We observed 0.4 γH2AX foci per nucleus in primary human melanocytes. In contrast, in four melanoma cell lines, we detected 7-17 γH2AX foci per nucleus, a 17-42 times increase in the basal level of γH2AX foci in melanoma cells relative to melanocytes (MC). Thus, untreated melanoma cells express significantly greater numbers of γH2AX foci than do untreated MC. Detection and rejoining of ionizing radiation-induced DNA dsb proceeded as rapidly in melanoma cells as in MC. Melanoma cells, however, reduced the number of radiation-induced γH2AX foci down only to pre-irradiation levels. Co-localization of the majority of γH2AX foci with ataxia telangiectasia mutated, BRCA1, 53BP1, and Nbs1 foci in untreated melanoma cells indicated that the additional foci in melanoma cells were associated with a DNA change that the cells interpret as DNA dsb. Co-localization of γH2AX foci with the telomere replication factor 1 protein in untreated melanoma cells indicates that the additional foci in untreated melanoma cells are associated with dysfunctional telomeres that induce a DNA damage stress response.
AB - When human cells sustain a DNA double-strand break (dsb), histone H2AX in chromatin surrounding the DNA break is phosphorylated, marking repair foci. The number of phosphorylated histone H2AX (γH2AX) foci approximates the number of dsb present in the cell's nuclear DNA. We observed 0.4 γH2AX foci per nucleus in primary human melanocytes. In contrast, in four melanoma cell lines, we detected 7-17 γH2AX foci per nucleus, a 17-42 times increase in the basal level of γH2AX foci in melanoma cells relative to melanocytes (MC). Thus, untreated melanoma cells express significantly greater numbers of γH2AX foci than do untreated MC. Detection and rejoining of ionizing radiation-induced DNA dsb proceeded as rapidly in melanoma cells as in MC. Melanoma cells, however, reduced the number of radiation-induced γH2AX foci down only to pre-irradiation levels. Co-localization of the majority of γH2AX foci with ataxia telangiectasia mutated, BRCA1, 53BP1, and Nbs1 foci in untreated melanoma cells indicated that the additional foci in melanoma cells were associated with a DNA change that the cells interpret as DNA dsb. Co-localization of γH2AX foci with the telomere replication factor 1 protein in untreated melanoma cells indicates that the additional foci in untreated melanoma cells are associated with dysfunctional telomeres that induce a DNA damage stress response.
KW - Chromosome instability
KW - Melanocyte
KW - Micronuclei
KW - Telomere dysfunction
KW - γH2AX foci
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U2 - 10.1111/j.0022-202X.2005.23674.x
DO - 10.1111/j.0022-202X.2005.23674.x
M3 - Article
C2 - 15816840
AN - SCOPUS:16844370883
SN - 0022-202X
VL - 124
SP - 807
EP - 817
JO - Journal of Investigative Dermatology
JF - Journal of Investigative Dermatology
IS - 4
ER -