Recently, microRNAs (miRNAs) have reported to participate in multiple biological processes. However, the effects of miR-495 on gastric cancer (GC) remain unclear. The purpose of this study is to explore the functions of miR-495 in GC cells proliferation, metastasis and apoptosis. SGC-7901 and BGC-823 were transfected with miR-495 mimic, miR-495 inhibitor and negative controls (mimic control and inhibitor control). The expressions of miR-495, cell viability, migration, apoptosis and apoptosis-related factors were examined by qRT-PCR, trypan blue staining, Transwell, flow cytometry and western blot, respectively. Simultaneously, the key factors expressions of EMT were detected by qRT-PCR again. The direct target gene of miR-495 was confirmed by dual luciferase assay. Additionally, sh-Twist1, pc-Twist1 and corresponding control were also transfected into SGC-7901 and BGC-823 cells, and then the protein levels of EMT-associated factors were detected by western blot. MiR-495 was down-regulated in GC cells. MiR-495 expression level was effectively overexpressed or suppressed in SGC-7901 and BGC-823 cells. Overexpression of miR-495 significantly decreased cell viability, migration and increased apoptosis as well as inhibited EMT process. Suppression of miR-495 showed contrary results. Twist1 was clarified as a target gene of miR-495, and Twist1 silencing obviously reduced the promotion effect of miR-495 suppression on these biological processes. Besides, Twist1 silencing significantly blocked EMT signal pathway in both SGC-7901 and BGC-823 cells. MiR-495 inhibited proliferation, metastasis and promoted apoptosis by targeting Twist1 in GC cells. These data indicated that miR-495 might be a novel anti-tumor factor of GC and provide a new method for treatment of GC.