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# The Role of Glutathione in the Aerobic Radioresponse: I. Sensitization and Recovery in the Absence of Intracellular Glutathione

Edward P. Clark, Edward R. Epp, Michele Morse-Gaudio and John E. Biaglow
Vol. 108, No. 3 (Dec., 1986), pp. 238-250
DOI: 10.2307/3576912
Stable URL: http://www.jstor.org/stable/3576912
Page Count: 13
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## Abstract

The effect of changes in both the intracellular glutathione (GSH) concentration and the concentration of extracellular reducing equivalents on the aerobic radiosensitization was studied in three cell lines: CHO-10B4, V79, and A549. Intracellular GSH was metabolically depleted after the inhibition of GSH synthesis by buthionine sulfoximine (BSO), while the extracellular environment was controlled through the replacement of growth medium with a thiol-free salt solution and in some experiments by the exogenous addition of either GSH or GSSG. Each of the cell lines examined exhibited an enhanced aerobic radioresponse when the intracellular GSH was extensively depleted (GSH < 1 nmol ${\rm GSH}/10^{6}\ \text{cells}$ after 1.0 mM BSO/24 h treatment) and the complexity of the extracellular milieu decreased. Although the addition of oxidized glutathione (5 mM GSSG/30 min) to cells prior to irradiation was without effect, much or all of the induced radiosensitivity was overcome by the addition of reduced glutathione (5 mM GSH/15 min). However, the observation that the exogenous GSH addition restores the control radio-response without increasing the intracellular GSH concentration was entirely unexpected. These results suggest that a number of factors exert an influence on the extent of GSH depletion and determine the extent of aerobic radiosensitization. Furthermore, the interaction of exogenous GSH with-but without penetrating-the cell membrane is sufficient to result in radio-recovery.

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