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Novel estradiol analogue induces apoptosis and autophagy in esophageal carcinoma cells

Abstract

Cancer is the second leading cause of death in South Africa. The critical role that microtubules play in cell division makes them an ideal target for the development of chemotherapeutic drugs that prevent the hyperproliferation of cancer cells. The new in silico-designed estradiol analogue 2-ethyl-3-O-sulfamoylestra-1,3,5(10)16-tetraene (ESE-16) was investigated in terms of its in vitro antiproliferative effects on the esophageal carcinoma SNO cell line at a concentration of 0.18 μM and an exposure time of 24 h. Polarization-optical differential interference contrast and triple fluorescent staining (propidium iodide, Hoechst 33342 and acridine orange) revealed a decrease in cell density, metaphase arrest, and the occurrence of apoptotic bodies in the ESE-16-treated cells when compared to relevant controls. Treated cells also showed an increase in the presence of acidic vacuoles and lysosomes, suggesting the occurrence of autophagic processes. Cell death via autophagy was confirmed using the Cyto-ID autophagy detection kit and the aggresome detection assay. Results showed an increase in autophagic vacuole and aggresome formation in ESE-16 treated cells, confirming the induction of cell death via autophagy. Cell cycle progression demonstrated an increase in the sub-G1 fraction (indicative of the presence of apoptosis). In addition, a reduction in mitochondrial membrane potential was also observed, which suggests the involvement of apoptotic cell death induced by ESE-16 via the intrinsic apoptotic pathway. In this study, it was demonstrated that ESE-16 induces cell death via both autophagy and apoptosis in esophageal carcinoma cells. This study paves the way for future investigation into the role of ESE-16 in ex vivo and in vivo studies as a possible anticancer agent.

Abbreviations

2ME:

2-methoxyestradiol

2-MeOE2bisMATE:

2-methoxyestradiolbis-sulfamate

AAF:

aggresome activity factor

AIF:

apoptosis inducing factor

AO:

acridine orange

Apaf-1:

apoptosis protease-activating factor

C9:

2-ethyl-3-Osulfamoylestra-1,3,5(10)-tetraen-3-ol-17-one

CAII:

carbonic anhydrase II

CAIX:

carbonic anhydrase IX

DMEM:

Dulbecco’s modified Eagle’s medium

DMSO:

dimethyl sulfoxide

EC:

esophageal cancer

ER:

estrogen receptor

ESE-16:

2-ethyl-3-O-sulfamoyl-estra-1,3,5(10)16-tetraene

FACS:

fluorescence-activated cell sorting

FB1:

fumonisin B1

HO:

Hoechst 33342

MFI:

mean fluorescent intensity

MOMP:

mitochondrial outer membrane permeabilization

PBS:

phosphate buffer saline

PE:

phosphatidylethanolamine

PI:

propidium iodide

PlasDIC:

polarization-optical transmitted light differential interference contrast microscopy

STS:

steroid sulfatase

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Correspondence to Annie Joubert.

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Wolmarans, E., Mqoco, T.V., Stander, A. et al. Novel estradiol analogue induces apoptosis and autophagy in esophageal carcinoma cells. Cell Mol Biol Lett 19, 98–115 (2014). https://doi.org/10.2478/s11658-014-0183-7

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  • DOI: https://doi.org/10.2478/s11658-014-0183-7

Keywords

  • Antiproliferative
  • 2-ethyl-3-O-sulfamoyl-estra-1,3,5(10)16-tetraene
  • ESE-16
  • Esophageal carcinoma
  • In vitro
  • Apoptosis
  • Autophagy