Increasing evidence has suggested the key roles of miRNAs in the initiation and progression of human cancers. miR-383 was downregulated and played a suppressive role in a variety of cancers; however, the function of miR-383 in gastric cancer remains unclear. In this study, we found that the expression of miR-383 was significantly reduced in gastric cancer tissues and correlated with the advanced progression of these cancer patients. Functional analysis showed that overexpression of miR-383 inhibited the proliferation and upregulated the apoptosis of gastric cancer cells. Furthermore, cyclin E2 was predicted as the target of miR-383 using the bioinformatics database. miR-383 bound the 3'-untranslated region of cyclin E2 and decreased the expression of cyclin E2 in gastric cancer cells. Upregulation of cyclin E2 was observed in gastric cancer tissues compared with the normal controls. Highly expressed cyclin E2 was inversely correlated with the level of miR-383 in gastric cancer tissues. Consistent with the decreased expression of cyclin E2 with miR-383, transfection of miR-383 induced cell cycle arrest at G1 phase in gastric cancer cells. Restoration of cyclin E2 significantly reversed the inhibitory effect of miR-183 on gastric cancer cell proliferation. Collectively, our results characterized the suppressive role of miR-383 in gastric cancer partially through targeting cyclin E2.