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The in vitro effects of 2-methoxyestradiol-bis-sulphamate on cell numbers, membrane integrity and cell morphology, and the possible induction of apoptosis and autophagy in a non-tumorigenic breast epithelial cell line

Cellular & Molecular Biology Letters volume 15, pages 564–581 (2010)Cite this article

Abstract

2-methoxyestradiol (2ME2) exerts estrogen receptor-independent anti-proliferative, anti-angiogenic and anti-tumor activity in vitro and in vivo. Due to its low bioavailability and rapid metabolic degradation, several analogues have been developed in recent years. 2-methoxyestradiol-bis-sulphamate (2-MeOE2bisMATE) is a bis-sulphamoylated derivative of 2ME2 with anti-proliferative activity. The aim of this study was to investigate cell signaling events induced by 2-MeOE2bisMATE in a non-tumorigenic cell line (MCF-12A) by analysing its influence on cell number, morphology and membrane integrity, and the possible induction of apoptosis and autophagy. Dose- and time-dependent studies revealed that 48 h exposure to 2-MeOE2bisMATE (0.4 μM) resulted in a decrease in cell numbers to 79%. A slight increase in the level of lactate dehydrogenase production was observed in the 2-MeOE2bisMATE-treated cells. Morphological studies revealed an increase in the number of cells in metaphase. Hallmarks of apoptosis were also found, namely nuclear fragmentation and apoptotic bodies. In addition, increased lysosomal staining was observed via fluorescent microscopy, suggesting the induction of another type of cell death, namely autophagy. Since 2-MeOE2bisMATE is regarded as a potential anti-cancer agent, it is also imperative to investigate the susceptibility of non-tumorigenic cells to its influence. The data generated from this study contributes to the understanding of the action that 2-MeOE2bisMATE exerts on the non-tumorigenic MCF-12A breast epithelial cell line.

Abbreviations

2ME2:

2-methoxyestradiol

2-MeOE2bisMATE:

2-methoxyestradiol-bis-sulphamate

adiol:

androstenediol

CA II:

carbonic anhydrase II

DHEA:

dehydroepiandrostene

DMSO:

dimethyl sulphoxide

LDH:

lactate dehydrogenase

PBS:

phosphate buffered saline

PlasDIC:

optical transmitted light differential interference contrast

STS:

steroid sulphatase

TEM:

transmission electron microscopy

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Authors and Affiliations

  1. Department of Physiology, University of Pretoria, Pretoria, P.O. Box 2034, Pretoria, 0001, South Africa

    Michelle H. Visagie & Annie M. Joubert

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  1. Michelle H. Visagie
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  2. Annie M. Joubert
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Correspondence to Annie M. Joubert.

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Visagie, M.H., Joubert, A.M. The in vitro effects of 2-methoxyestradiol-bis-sulphamate on cell numbers, membrane integrity and cell morphology, and the possible induction of apoptosis and autophagy in a non-tumorigenic breast epithelial cell line. Cell Mol Biol Lett 15, 564–581 (2010). https://doi.org/10.2478/s11658-010-0030-4

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Cellular & Molecular Biology Letters

ISSN: 1689-1392

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