Sufentanil is a new kind of opioid analgesic and acts on μ opioid receptor. In this study, we aim to investigate the effects of sufentanil on gastric cancer cell line SGC-7901, after being exposed to different concentrations of sufentanil. Gastric cancer cells were exposed to sufentanil for a predetermined time at concentrations of 0, 0.5, 5, 50 and 500 nmol/l, respectively. Cell viability at different time points after exposure to sufentanil was tested by CCK-8 assay. FDA-PI staining was used to observe membrane integrity of gastric cancer SGC7901 cells. The apoptosis of gastric cancer cells was analyzed by Annexin V-FITC/PI Flow Cytometry and the changes of the cell cycle was determined by a detection kit. As a result, cell viability decreased in a dose- and time-dependent manner. Furthermore, with the concentration of sufentanil increased, the proportion of dead and apoptotic SGC-7901 cells increased, and more cells were arrested in G2/M phase. In a word, sufentanil can inhibit the cell viability and induce the apoptosis of gastric cancer SGC-7901 cells in vitro.