Objective: GLP-1 exerts anti-apoptotic properties in pancreatic β-cell in type 2 diabetes mellitus. A main cause of the apoptosis is glucotoxiciy. However, the anti-apoptotic mechanism of GLP-1 and the duration for which GLP-1 should stimulate β-cells have not been fully determined. This study was to elucidate whether a short-term, physiological treatment with GLP-1 can be effective against β-cell apoptosis in response to high glucose, and its molecular mechanisms. Methods: INS-1 cells were cultured in RPMI-1640 and the common bile duct was cannulated and perfused through the pancreatic duct with HBSS containing collagenase (1 mg/mL, Collagenase Type V) to obtain the isolated islets. The anti-apoptosis effect of GLP-1 in INS-1 cells was investigated using a live cell movie analyser and cell viability was measured using the MTT assay. After the INS-1 cells were transfected with scrambled siRNA (scRNA), siRaptor or siRictor for 48 h, they were treated with GLP-1 for 30 min. After that the insulin secreted into the medium was collected and measured using an insulin enzyme immunoassay kit. Finally, the insulin level was standardized to the total protein content measured using the bicinchoninic acid assay Results: The protective effects of GLP-1 were related with PI3K/AKT S473 phosphorylation. Indeed, the increase in AKT S473 phosphorylation led to suppression of FoxO-1. Moreover, GLP-1-induced AKT S473 activation and FoxO-1 suppression were abolished by the selective inactivation of mTORC2 using siRNA directed towards the rapamycin-insensitive companion of the target of rapamycin. The protection effect of GLP-1 in β-cell apoptosis via glucotoxicity was also abolished by the selective inactivation of mTORC2. This report provides evidence that physiological short-term treatment with GLP-1 is sufficient to protect against glucotoxicity-induced β-cell apoptosis. Conclusion: The effect of GLP-1 may be mediated by FoxO-1 suppression through the PI3K/mTORC2/AKT S473 phosphorylation signalling pathways.