MicroRNA (miR)-185, which has been reported to be abnormally expressed in some types of cancer, exerts significant effects on the proliferation, apoptosis, drug resistance and metastasis of cancer cells. The present study aimed to explore the effects and underlying molecular mechanisms of miR-185 upregulation on the apoptosis of gastric cancer (GC) cells. Quantitative polymerase chain reaction (qPCR) and western blotting were used to detect the expression levels of miR-185 in GC and adjacent normal tissues. In addition, miR-185 expression was detected in the following GC cell lines: MKN74, SGC7901, BGC823, MGC803, as well as in the gastric epithelial cell line GES-1. Subsequently, miR-185 mimics were transfected into MGC803 cells. Post-transfection, the following experiments were conducted: MTT assay was applied to test cell viability; flow cytometry (FCM) was used to determine the apoptotic rate of the cells; and qPCR and western blotting were conducted to detect the expression levels of the following apoptosis-associated factors: B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), survivin, X-linked inhibitor of apoptosis protein (XIAP), livin, caspase-3 and caspase-8. The results demonstrated that miR-185 was downregulated in GC tissues compared with the adjacent tissues. In cell lines, miR-185 expression was higher in GES-1 cells compared with in the GC cell lines; in the 4 GC cell lines, the strongest miR-185 expression was in MKN74 cells, followed by SGC7901 and BGC823 cells, and the weakest was in MGC803 cells (P<0.05). Expression of miR-185 was associated with tumor size, differentiation and lymphatic metastasis. Post-transfection with miR-185 mimics, miR-185 expression was significantly increased in a time- and concentration-dependent manner. MGC803 cell viability was significantly decreased following miR-185 mimics transfection. The results of FCM demonstrated that post-transfection with miR-185 mimics, the apoptotic rate of MGC803 cells was significantly increased. Post-transfection with miR-185 mimics, the expression levels of Bcl-2, survivin and XIAP were significantly decreased in MGC803 cells, whereas the expression levels of Bax and livin were not altered, and caspase-3 and caspase-8 expression was significantly increased. Spectrophotometry indicated that caspase-3 and caspase-8 activity was significantly increased in MGC803 cells following transfection with miR-185 mimics. In conclusion, the present study suggested that miR-185 upregulation in GC cells may promote apoptosis of tumor cells via gene expression regulation.