Despite its limited success, 5-fluorouracil (5-FU) remains the primary chemotherapy agent for the treatment of esophageal cancer. Quercetin has been demonstrated to inhibit the growth of transformed cells. The present study was conducted to examine whether quercetin combined with conventional chemotherapeutic agents would improve the therapeutic strategy for esophageal cancer. In this study, an MTT assay was used to determine the effects of quercetin on the proliferation of EC9706 and Eca109 cells. Annexin V-FITC/propidium iodide (PI)-stained fluorescence-activated cell sorter (FACS) analysis was used to detect the apoptotic fraction of treated cells, and western blot analysis was used to examine the protein levels. The results of our study demonstrated that quercetin in combination with 5-FU significantly inhibited growth (P<0.05) and stimulated apoptosis (P<0.005) in EC9706 and Eca109 esophageal cancer cells compared with quercetin or 5-FU alone. These changes were associated with the decreased expression of a phosphorylated inhibitory molecule of NF-κB (pIκBα), which was activated by exposure to 5-FU alone. We suggest that inclusion of quercetin to the conventional chemotherapeutic agent 5-FU may be an effective therapeutic strategy for esophageal cancer.