Esophageal cancer is a common malignant tumor occurring in human esophageal epithelial tissue. The primary purpose of this paper was to define the effects of ${eta}$-carotene and 1,25-dihydroxyvitamin $D_3$, alone and in combination, on cell proliferation, cell cycle and apoptosis of human esophageal cancer EC9706 cells. Treatment with different concentrations of ${eta}$-carotene and/or 1,25-dihydroxyvitamin $D_3$. MTT assay showed that ${eta}$-carotene and 1,25-dihydroxyvitamin $D_3$ significantly inhibited proliferation of EC9706 cells in a dose- and time-dependent manner. Further studies also demonstrated that ${eta}$-carotene alone or 1,25-dihydroxyvitamin $D_3$ alone caused a marked increase on the induction of apoptosis in EC9706 cells. The percentage of G0/G1-phase cells significantly increased on addition of 1,25-dihydroxyvitamin $D_3$ alone, but there were no significant changes with ${eta}$-carotene alone. These two agents in combination synergistically inhibited cell growth and induced apoptosis. Therefore, our results indicate that ${eta}$-carotene and 1,25-dihydroxyvitamin $D_3$ in combination may provide a novel strategy for preventing and treating esophageal cancer.