2-Methoxyestradiol-bis-sulphamate (2-MeOE2bisMATE) is a bis-sulphamoylated analogue of a biological estradiol metabolite, 2-methoxyestradiol (2ME2) with potential as an anticancer drug. The aim of this in vitro study was to evaluate the effects of 2-MeOE2bisMATE on alpha-tubulin structure, as well as its potential to induce apoptosis in an esophageal carcinoma (SNO) cell line using confocal microscopy, flow cytometry and spectrophotometry. 2-MeOE2bisMATE disrupted the microtubule network of SNO cells, arrested cells in metaphase and induced apoptosis. An increase in the number of cells present in sub-G1, mitochondrial membrane potential reduction and increased caspase 6 activity were observed. This in vitro study demonstrated new insights on the action mechanism of 2 MeOE2bisMATE in esophageal carcinoma (SNO) cells, since these activities have not been studied in esophageal carcinoma cells up to date. Future studies are warranted to further determine which gene and protein expression changes are induced by 2-MeOE2bisMATE in SNO cells.