The present study aimed to investigate the role of the extracellular signal-regulated kinase (ERK)1/2 signal transduction pathway in glucose‑stimulated insulin secretion in β‑TC6 mouse pancreatic cells. Insulin production by β‑TC6 cells was stimulated with various concentrations of glucose, which was dose-dependently inhibited by mitogen‑activated protein kinase inhibitor PD98059, as indicated by a radioimmunoassay. Furthermore, glucose stimulation enhanced the phosphorylation of ERK1/2, which was dose-dependently inhibited by PD98059, as indicated by western blot analysis. These results indicated that the activation of the ERK1/2 signal transduction pathway may have an important role in glucose‑stimulated insulin secretion in β‑TC6 cells.