Objective:To establish mouse model of PP2AC gene specifical inactivation in pancreatic cells and investigate preliminary animal phenotype. Methods: The Ins-2 transgenic mice in which Cre enzyme was exclusively detected in pancreatic-cells bred with PP2ACαflox/flox mice to obtain PP2ACαflox/+:Ins-2 mice, then bred with the PP2ACαflox/flox mice again to obtain PP2ACαflox/flox:Ins-2 mice (KO mice). We identified the 2nd exon of PP2ACα knockdown by PCR and Western Blot. Interperitoneal glucose tolerance test (IPGTT) was performed in transgenic mice at 4 months, and PP2ACαflox/flox mice were employed with control. Results: PP2ACα transcripts were shorter in KO mice than those in control mice. The protein level of PP2AC expression in KO mice was significantly lower than that of control mice (P < 0.05). The results of IPGTT indicated that the blood glucose levels at 30 min, 60 min and 120 min were significantly higher than those of controls (P < 0.05). Conclusion: Specifical inactivation of PP2AC in pancreatic β-cells mouse model is successfully established, and glucose tolerance impairs at 4 months in PP2ACαflox/flox:Ins2-Cre mice.