Molecular crosstalk between apoptosis and autophagy induced by a novel 2-methoxyestradiol analogue in cervical adenocarcinoma cells

Cancer Cell International

Annie M Joubert²  Laurence Lafanechère¹  Elsie M Nolte²  Anne E Theron² 

[1] Department of Cellular Differentiation and Transformation, Team# 03: Polarity, Development and Cancer, Université Joseph Fourier, Albert Bonniot Institute, CRI INSERM/UJF U823, Grenoble, France;Department of Physiology, Faculty of Health Sciences, University of Pretoria, Private Bag X323, Arcadia, 0007 Gauteng, Pretoria, South Africa

关键词: 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene;    Microtubules;    Mitotic block;    Crosstalk;    Caspase;    Autophagy;    Apoptosis;    Analogue;    ESE-16;    2-methoxyestradiol;   

Others  :  792967
DOI  :  10.1186/1475-2867-13-87

 received in 2013-05-16, accepted in 2013-08-23,  发布年份 2013

【 摘 要 】

Background

2-Methoxyestradiol has been shown to induce both autophagy and apoptosis in various carcinogenic cell lines. Although a promising anti-cancer agent, it has poor bioavailability and rapid in vivo metabolism which decreases its efficiency. In order to improve 2-methoxyestradiol’s anti-proliferative properties, a novel 2-methoxyestradiol analogue, 2-ethyl-3-O-sulphamoyl-estra-1,3,5 (10)16-tetraene (ESE-16), was previously in silico-designed in our laboratory. This study investigated ESE-16 for its anti-proliferative potential on a cervical adenocarcinoma cell (HeLa) cell line. Additionally, the possible intracellular crosstalk mechanisms between the two types of cell death were investigated.

Methods and results

HeLa cells exposed to 0.5 μM ESE-16 for 24 hours showed morphological evidence of both apoptotic and autophagic death pathways as assessed by polarization-optical transmitted light differential interference contrast microscopy, fluorescent microscopy and transmission electron microscopy. Flow cytometric cyclin B1 quantification revealed induction of programmed cell death after halting cell cycle progression in metaphase. Confocal microscopy demonstrated that ESE-16 caused microtubule fragmentation. Flow cytometric analysis of cell cycle progression and phosphatidylserine flip determination confirmed induction of apoptosis. Moreover, an increase in aggresome formation and microtubule-associated protein light chain, LC3, was demonstrated indicative of autophagy. Both caspase 8 and 3 were upregulated in a spectrophotometric analysis, indicating the involvement of the extrinsic pathway of apoptotic induction.

Conclusions

We conclude that the novel in silico-designed compound, ESE-16, exerts its anti-proliferative effect on the tumorigenic human epithelial cervical (HeLa) cells by sequentially targeting microtubule integrity, resulting in a metaphase block, causing induction of both autophagic and apoptotic cell death via a crosstalk mechanism that involves the extrinsic pathway. Future investigations will expand on signal transduction pathways involved in both apoptosis and autophagy for assessment of ESE-16 effects on microtubule dynamic instability parameters.

【 授权许可】

   
2013 Theron et al.; licensee BioMed Central Ltd.

【 图 表 】

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