Gastric cancer is the second most common cause of cancer-related death worldwide. This study was designed to examine the role of CUGBP1 in cell growth via an RNA interference (RNAi) lentivirus system in gastric cancer cells in vitro. The expression of CUGBP1 was much stronger in gastric cancer tissues than that in adjacent normal tissues. The lentivirus-mediated knockdown of CUGBP1 resulted in a significant reduction of CUGBP1 expression in MGC-803 gastric cancer cells. The cell viability was remarkably decreased by 50 % after 5 days of infection, as determined by MTT assay. Moreover, the size and the number of colonies formed in MGC-803 cells were markedly reduced in the absence of CUGBP1. Furthermore, the silencing of CUGBP1 downregulated the expression levels of cyclin B1 and cyclin D1, which are involved in cell cycle control. These results clearly indicated that CUGBP1 is essential for the growth of gastric cancer cells. Therefore, silencing of CUGBP1 by RNAi could be developed as a promising therapeutic approach for gastric cancer.