BACKGROUND: Anticancer research resulted in the discovery of a promising antimitotic metabolite,
2-methoxyestradiol. 2-Methoxyestradiol-bis-sulphamate, a bis-sulphamoylated analogue exerts antiproliferative- and
antimitotic activity. Investigating the anticancer potential of 2-methoxyestradiol-bis-sulphamate requires
demonstrating the influence of 2-methoxyestradiol-bis-sulphamate on non-tumorigenic cells. This project focused
on the in vitro effects of 2-methoxyestradiol-bis-sulphamate on the non-tumorigenic MCF-12A breast epithelial cell line.
METHODS The in vitro influence of 2-methoxyestradiol-bis-sulphamate was investigated on cell cycle progression, possible induction of apoptosis and autophagy and reactive oxygen species generation. Cell cycle progression was done using flow cytometry in conjunction with ethanol fixation and propidium iodide staining. Displaying effects
on the mitochondrial membrane potential was achieved utilizing flow cytometry and the MitoCapture TM Mitochondrial apoptosis detection kit. Autophagy detection was done by means of flow cytometry and anti-LC3B
conjugated to DyLight 488. Reactive oxygen species generation was conducted employing flow cytometry and
2,7-dichlorofluorescein diacetate and hydroethidine.
RESULTS: This study demonstrated that 2-methoxyestradiol-bis-sulphamate did not affect cell cycle progression or
reactive oxygen species in a statistically significant manner in the non-tumorigenic MCF-12A cell line. In addition,
2-methoxyestradiol-bis-sulphamate did not statistically significantly induce apoptosis or autophagy.
CONCLUSION: Reports indicate that 2-methoxyestradiol-bis-sulphamate induces apoptosis and autophagy in several
tumorigenic cell lines. The anticancer ability of 2-methoxyestradiol-bis-sulphamate is due to its antimitotic activity.
However, this study demonstrates the promising notion that 2-methoxyestradiol-bis-sulphamate does not affect the
non-tumorigenic MCF-12A cells. This project contributes to the embedded scientific knowledge regarding the
differential death mechanisms used by 2-methoxyestradiol-bis-sulphamate on tumorigenic and non-tumorigenic