O-GlcNAcylation mediates H2O2-induced apoptosis through regulation of STAT3 and FOXO1

The O-linked-β-N-acetylglucosamine (O-GlcNAc) glycosylation (O-GlcNAcylation) is a critical post-translational modification that couples the external stimuli to intracellular signal transduction networks. However, the critical protein targets of O-GlcNAcylation in oxidative stress-induced Apoptosis remain to be elucidated. Here, we show that treatment with H₂O₂ inhibited O-GlcNAcylation, impaired cell viability, increased the cleaved Caspase 3 and accelerated Apoptosis of neuroblastoma N2a cells. The O-GlcNAc transferase (OGT) inhibitor OSMI-1 or the O-GlcNAcase (OGA) inhibitor Thiamet-G enhanced or inhibited H₂O₂-induced Apoptosis, respectively. The total and phosphorylated protein levels, as well as the promoter activities of signal transducer and activator of transcription factor 3 (STAT3) and Forkhead box protein O 1 (FOXO1) were suppressed by OSMI-1. In contrast, overexpressing OGT or treating with Thiamet-G increased the total protein levels of STAT3 and FOXO1. Overexpression of STAT3 or FOXO1 abolished OSMI-1-induced Apoptosis. Whereas the anti-apoptotic effect of OGT and Thiamet-G in H₂O₂-treated cells was abolished by either downregulating the expression or activity of endogenous STAT3 or FOXO1. These results suggest that STAT3 or FOXO1 are the potential targets of O-GlcNAcylation involved in the H₂O₂-induced Apoptosis of N2a cells.

FOXO1; O-GlcNAcylation; STAT3; apoptosis; oxidative stress.