2-Methoxyestradiol (2ME2) is a natural metabolite of 17-β-estradiol exerting both antiproliferative and antiangiogenic characteristics. Due to limited bioavailability, analogues of 2ME2 were designed, synthesized and evaluated for improved in vitro antiproliferative activity as well as bioavailability. 2-Methoxyestradiol-bis-sulphamate (2-MeOE2bisMATE) is an analogue of 2ME2 which was produced by sulphamoylation of the two hydroxyl groups on carbons 3 and 17. The aim of this in vitro study was to evaluate the influence of 2- MeOE2bisMATE on morphology, apoptosis and autophagy induction in an oesophageal carcinoma (SNO) cell line by means of transmission electron microscopy (TEM) and flow cytometry (cyto-ID and LC3 autophagy detection assays). In 2-MeOE2bisMATE-treated cells, features of both apoptosis and autophagy such as the presence of apoptotic bodies and autophagy vesicles were observed using the transmission electron microscopy. A significant increase in the quantity of autophagy vesicles and LC3 levels were observed in 2- MEOE2bisMATE-treated cells. This in vitro study demonstrates that 2-MeOE2bisMATE induces both apoptosis and autophagy which was revealed by the increase in apoptosis- and autophagy-associated morphology, as well as by an increase in the measure of autophagy vesicles and LC3 levels in SNO cells. From data observed from this study, 2-MeOE2bisMATE is shown to be a potential anticancer agent; further in vitro research is, however, warranted.