Objective To study the acute cytotoxicity of four kinds of Alternaria toxins alternaiol (AOH), alternariol monomethyl ether (AME), tenuazonic acid (TeA) and tentoxin (TEN) on human esophageal epithelium cell Het-1 A in vitro. Methods Het-1 A was treated with different concentrations of 4 kinds of Alternaria toxins. The effects of 4 kinds of Alternaria toxins on cell proliferations, cell apoptosis, cell cycle distribution and cysteinyl aspartate specific proteinase-3 (capase-3) activity were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide(MTT), annexin V-fluorescein isothiocyanate/propidium iodide(Annexin V-FITC/PI)double staining method, PI single staining method and spectrophotometry, respectively. Results The half maximal inhibitory concentration (IC₅₀) of AOH, AME, TeA and TEN on Het-1 A was 54.31,3.38,1.91 and 141.96 μmol/L, respectively. The 4 kinds of Alternaria toxins could induce the apoptosis of Het-1 A and also could affect the cell cycle distribution through increasing the percent of G₂-M phase. AOH and AME could enhance the activity of caspase-3 in a positive dose-dependent manner to induce the apoptosis of Het-1 A. Conclusion AOH, AME, TeA and TEN could cause acute cytotoxicity on Het-1 A through inhibiting cell proliferation, inducing cell apoptosis and G₂-M cell cycle arrest etc.