EXOGENOUS GLUTATHIONE ATTENUATES THE ANTIPROLIFERATIVE EFFECT OF BUTHIONINE SULFOXIMINE

Buthionine sulfoximine (BSO) inhibits proliferation of human lung carcinoma A549 cells in a manner that does not correlate with intracellular glutathione (GSH) depletion, nor does it reflect overt toxic effects of BSO. However, BSO inhibits uptake by A549 cells of cystine, which is an essential amino acid for cell growth in culture. Thus, it is hypothesized that inhibition of cellular cystine uptake is, or is partially, responsible for the antiproliferative effect of BSO. It has been shown that the gamma-glutamyl amino acid transport system plays a role in cystine transport across cell membranes. This transport system requires extracellular GSH for its operation. BSO, by inhibiting intracellular GSH synthesis, would reduce GSH export and decrease extracellular GSH levels. Therefore, the present study was undertaken to examine the effect of exogenously added GSH on BSO inhibition of cellular cystine uptake and its relationship to the antagonistic effect of GSH on BSO antiproliferation. A549 cells were treated with 10 mM BSO and exogenous GSH was added to these BSO-treated cultures. Effects of exogenous GSH on BSO antiproliferation and cellular GSH depletion were determined simultaneously as a function of time. The effect of GSH on BSO inhibition of cystine accumulation was measured using [S-35]cystine. The results obtained demonstrate that exogenously added GSH partially overcame BSO antiproliferation. The GSH antagonistic effect did not correlate with repletion of intracellular GSH, but it did correlate with recovery of BSO-inhibited cystine accumulation. Exogenous GSH also enhanced proliferation of non-BSO treated cells at concentrations below 1.0 mM. The results of this study suggest that BSO inhibition of cystine uptake may represent one mechanism by which BSO exerts its antiproliferative effect. The antagonistic effect of exogenous GSH on BSO antiproliferation may result from recovery of BSO-inhibited cystine uptake, although other mechanisms responsible for the GSH antagonistic effect may also exist.