Type 2 diabetes is a metabolic disorder characterized by a defect in insulin secretion and a decrease of peripheral insulin sensitivity. Glucotoxicity, the detrimental effect of glucose, is involved in this disease progression, and has been shown to exert deleterious effects on various organs and cells, notably leading to impairment of insulin secretion by pancreatic β-cells. Mechanisms of glucose-induced β-cell failure are not entirely described and are fundamental to figure out how β-cells respond to increased glucose stimulation and finally undergo apoptosis. Therefore, the overall goal of this study was to monitor the early molecular mechanisms underlying β-cell dysfunction in conditions of chronic high glucose exposition.