2-Methoxyestradiol-bis-sulphamate refrains from inducing apoptosis and autophagy in a non-tumorigenic breast cell line

Michelle H Visagie,Anna M Joubert

doi:10.1186/1475-2867-12-37

Abstract

BackgroundAnticancer 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.MethodsThe 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.ResultsThis 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.ConclusionReports 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 cell lines.

Highlights

Researchers have identified a promising anticancer compound, 2-methoxyestradiol (2ME), that exerts antimitotic- and antiproliferative activity in vitro and in vivo [1,2]
Apoptosis was induced by 2-Methoxyesradiol-bis-sulphamate is (2MEBM) in MCF-7, prostate cancer cells (PC-3), human umbilical vein endothelial cells (HUVEC) and the human breast adenocarcinoma CAL51 cell line [13,14,15]
The Annexin V fluorescein isothiocyanate (FITC) kit, MitoCapture TM Mitochondrial apoptosis detection kit and a rabbit polyclonal anti-LC3B conjugated to DyLight 488 were purchased from BIOCOM biotech (Pty) Ltd. (Clubview, South Africa). 2,7-Dichlorofluorescein diacetate and hydroethidine was acquired from Sigma

Summary

Introduction

Researchers have identified a promising anticancer compound, 2-methoxyestradiol (2ME), that exerts antimitotic- and antiproliferative activity in vitro and in vivo [1,2]. 2-Methoxyesradiol-bis-sulphamate is (2MEBM) a bissulphamoylated derivative of 2ME that exerts antiproliferative- and anticancer activity with improved bioavailability and a superior pharmacokinetic profile [57]. The estrogen 3-O-sulphamates strongly inhibit carbonic anhydrase II which is responsible for the conversion of carbon dioxide and water to carbonic acid This interaction is most likely responsible for the high bioavailability of the sulphamoylated analogues as reversible uptake by red blood cells and interaction with carbonic anhydrase II ensures transiting the liver without undergoing first pass metabolism [1,17,18]. 2-Methoxyestradiol-bis-sulphamate, a bis-sulphamoylated analogue exerts antiproliferative- and antimitotic activity. This project focused on the in vitro effects of 2-methoxyestradiol-bis-sulphamate on the non-tumorigenic MCF-12A breast epithelial cell line

Methods

Results

Discussion

Conclusion