MicroRNAs (miRs) are considered to be effective, post-transcriptional regulators in the pathophysiology of type 2 diabetes (T2D) and promising treatment targets. However, the function of miR-141 remains to be elucidated. In the present study, upregulation of miR-141 was demonstrated in diabetic mice and elderly diabetic patients. Using reverse transcriptase-quantitative polymerase chain reaction, luciferase reporter assays and western blotting, forkhead box A2 (FOXA2) was identified as a direct target gene of miR-141. The potential role of miRNA-141 or FOXA2 was evaluated by overexpressing or silencing miR-141 or FOXA2, respectively. The increased expression of miR-141 resulted in impaired glucose-stimulated insulin secretion (GSIS) and INS-1 cell proliferation. In addition, miR-141 silencing in MIN6 pseudoislets or INS-1 cells led to reduced T2D-associated damage. Furthermore, the expression of miR-141 may be corrected by treatment with pioglitazone, which is widely used for insulin resistance therapy. The present study also demonstrated the mechanism by which miR-141 regulated GSIS and proliferation through FOXA2. Overexpression of FOXA2 in MIN6 pseudoislets increased the effect of the miR-141 inhibitor on GSIS. FOXA2 effectively reversed the effect of miR-141 overexpression on cell proliferation. In conclusion, the results of the present study indicate that the pioglitazone/miR-141/FOXA2 axis may represent a promising target mechanism for T2D treatment.