Radiosensitization of mouse skin by oxygen and depletion of glutathione.
OBJECTIVE
To determine the oxygen enhancement ratio (OER) and shape of the oxygen sensitization curve of mouse foot skin, the extent to which glutathione (GSH) depletion radiosensitized skin, and the dependence of such sensitization on the ambient oxygen tension.


METHODS
The feet of WHT mice were irradiated with single doses of 240 kVp x-rays while mice were exposed to carbogen or gases with oxygen/nitrogen mixtures containing 8-100% O2. The anoxic response was obtained by occluding the blood supply to the leg of anesthetized mice with a tourniquet, surrounding the foot with nitrogen, and allowing the mice to breathe 10% O2. Further experiments were performed to assess the efficacy of this method to obtain an anoxic response. Radiosensitivity of skin was assessed using the acute skin-reaction assay. Glutathione levels were modified using two schedules of DL-buthionine sulphoximine (BSO) and diethylmaleate (DEM), which were considered to produce extensive and intermediate levels of GSH depletion in the skin of the foot during irradiation.


RESULTS
Carbogen caused the greatest radiosensitization of skin, with a reproducible enhancement of 2.2 relative to the anoxic response. The OER of 2.2 is lower than other reports for mouse skin. This may indicate that the extremes of oxygenation were not produced, although there was no direct evidence for this. When skin radiosensitivity was plotted against the logarithm of the oxygen tension in the ambient gas, a sigmoid curve with a K value of 17-21% O2 in the ambient gas was obtained. Depletion of GSH caused minimal radiosensitization when skin was irradiated under anoxic or well-oxygenated conditions. Radiosensitization by GSH depletion was maximal at intermediate oxygen tensions of 10-21% O2 in the ambient gas. Increasing the extent of GSH depletion led to increasing radiosensitization, with sensitization enhancement ratios of 1.2 and 1.1, respectively, for extensive and intermediate levels of GSH depletion. In mice exposed to 100% O2, a significant component of skin radiosensitivity was due to diffusion of oxygen directly through the skin. Pentobarbitone anesthesia radiosensitized skin in mice exposed to 100% O2 by a factor of 1.2, but did not further sensitize skin in mice exposed to carbogen.


CONCLUSIONS
Glutathione levels and the local oxygen tension at the time of irradiation were important determinants of mouse foot skin radiosensitivity. The extent to which GSH levels altered the radiosensitivity of skin was critically dependent on the local oxygen tension. These results have significant implications for potential clinical application of GSH depletion.
Stevens
G
G
Gray Laboratory of the Cancer Research Campaign, Mount Vernon Hospital, Northwood, Middlesex, UK.
Joiner
M
M
Joiner
B
B
Johns
H
H
Denekamp
J
J
D000818:Animals; D019328:Buthionine Sulfoximine; D002245:Carbon Dioxide / Q000494:pharmacology*; D015687:Cell Hypoxia / Q000187:drug effects; D004307:Dose-Response Relationship, Radiation; D005260:Female; D005978:Glutathione / Q000172:deficiency* / Q000187:drug effects; D008297:Male; D008298:Maleates / Q000494:pharmacology; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008920:Misonidazole / Q000494:pharmacology*; D010100:Oxygen / Q000494:pharmacology*; D010101:Oxygen Consumption; D010313:Partial Pressure; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D012867:Skin / Q000098:blood supply / Q000378:metabolism / Q000528:radiation effects*; D014111:Tourniquets
D008298:Maleates; D011838:Radiation-Sensitizing Agents; D002245:Carbon Dioxide; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; C011700:carbogen; D008920:Misonidazole; C014476:diethyl maleate; D005978:Glutathione; D010100:Oxygen