Protection from radiation-induced DNA single-strand breaks by induction of nuclear metallothionein.
OBJECTIVE
To examine the extent to which nuclear metallothionein protects from radiation-induced DNA damage under aerobic and hypoxic conditions.


METHODS
A semiquantitative fluorescence image analysis method measured the nuclear content of metallothionein (MT) in ME180 and SiHa human squamous cervical carcinoma cell lines under normal growth conditions, and following MT induction by zinc. The extent of initial DNA damage following 60Co irradiation under aerobic and hypoxic conditions was assessed using the alkaline comet assay.


RESULTS
Provided that cells were maintained at 37 degrees C, most of the cellular content of MT was in the nucleus. Incubation at 4 degrees C caused the rapid translocation of MT from the nucleus into the cytoplasm in both cell lines, with no net loss of cellular MT. Baseline nuclear MT levels were about four times greater in ME180 cells, and were much more readily induced by treatment with 100 microM zinc acetate, compared with SiHa cells. Under aerobic conditions, MT induction by zinc resulted in no protection in either of the cell lines. Under hypoxic conditions, however, the number of DNA single-strand breaks in zinc-treated cells was reduced by approximately 40% in ME180, but not in SiHa cells, when compared with non-induced controls.


CONCLUSIONS
Nuclear MT can exert a significant level of protection from radiation by a mechanism that involves competition with oxygen for DNA radical sites and/or scavenging of free radicals. Because increased MT levels have been reported in hypoxic micro-regions of some solid tumours, this protective mechanism might have clinical relevance.
Vukovic
V
V
Ontario Cancer Institute-Princess Margaret Hospital, Department of Medical Biophysics, University of Toronto, Ontario, Canada.
Pheng
S R
SR
Stewart
A
A
Vik
C H
CH
Hedley
D W
DW
D000332:Aerobiosis; D015687:Cell Hypoxia; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D003545:Cysteine / Q000378:metabolism; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000528:radiation effects*; D005260:Female; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D008668:Metallothionein / Q000096:biosynthesis*; D011837:Radiation-Protective Agents / Q000378:metabolism; D013696:Temperature; D014407:Tumor Cells, Cultured; D015032:Zinc / Q000494:pharmacology
D004273:DNA, Neoplasm; D011837:Radiation-Protective Agents; D008668:Metallothionein; D005978:Glutathione; D015032:Zinc; D003545:Cysteine