MicroRNA-145 (miR-145) has been shown to regulate the development of different human cancer. However, the role of miR-145 via modulation of tuftelin 1 (TUFT1) expression has not been studied in gastric cancer. TUFT1The results showed that gastric cancer tissues and cell lines exhibit significant (P<0.05) downregulation of miR-145. Overexpression of miR-145 significantly (P<0.05) inhibited the viability and colony formation of the MGC-803 gastric cancer cells. Annexin V/PI staining revealed that miR-145 exerts its tumor-suppressive effects via induction of apoptosis. The apoptotic cell percentage increased from 5.75% in negative control to 22.95% in miR-145 overexpressing MG-803 cells. This was also accompanied by upregulation of Bax and downregulation of Bcl-2 expression. TargetScan analysis and the dual luciferase assay revealed TUFT1 as the functional target of miR-145. The expression of TUFT1 was significantly (P<0.05) upregulated in gastric cancer tissues and cell lines. However, overexpression of miR-145 causes inhibition of the TUFT1 expression. Silencing of TUFT1 mimicked the tumor-suppressive effects of miR-145. However, tuftelin 1 overexpression attenuated the tumor-suppressive effect of miR-145 in MGC-803 gastric cancer cells. Taken together, the results of the present study indicate that miR-145 targets TUFT1 at translational level to exert its tumor suppressive effects in gastric cancer.