Rapid induction of the intrinsic apoptotic pathway by L-glutamine starvation

While the amino acid L-glutamine is known to play a role in the survival of several cell types,the underlying molecular mechanisms are still poorly defined. We show in this report that L-glutamine starvation rapidly triggered apoptosis in Sp2/0-Ag14 hybridoma cells. This process involved the activation of both caspases-9 and -3, suggesting that L-glutamine deprivation initiated an intrinsic apoptotic pathway in Sp2/0-Ag14 cells. Supporting this idea, the cytosolic release of the mitochondrial proteins SMAC/DIABLO and cytochrome c (Cyt c) was observed, with an initial limited leakage occurring during the first 30 min Of L-glutamine deprivation, followed by a greater release after 60 min. The latter occurred simultaneously with the trans location of the pro-apoptotic protein Bax to the mitochondria. Finally, a decline in XIAP levels and the activation of caspases-3 and -9 were observed. Thus, L-glutamine deprivation of Sp2/0-Ag14 cells rapidly triggers intracellular events, which target the mitochondria, leading to the cytosolic release of apoptogenic factors, the activation of caspases-9 and -3, and the commitment to the death program. This work introduces the Sp2/ 0Ag14 hybridoma as a unique model for the study of the molecular events underlying the pro-survival function Of L-glutamine. (C) 2004 Wiley-Liss, Inc.