Abstract
We have recently reported that cytostatic concentrations of the microsomal antiestrogen-binding site (AEBS) ligands, such as PBPE (N-pyrrolidino-(phenylmethyphenoxy)-ethanamine,HCl) and tamoxifen, induced differentiation characteristics in breast cancer cells through the accumulation of post-lanosterol intermediates of cholesterol biosynthesis. We show here that exposure of MCF-7 (human breast adenocarcinoma cell line) cells to higher concentrations of AEBS ligands triggered active cell death and macroautophagy. Apoptosis was characterized by Annexin V binding, chromatin condensation, DNA laddering and disruption of the mitochondrial functions. We determined that cell death was sterol- and reactive oxygen species-dependent and was prevented by the antioxidant vitamin E. Macroautophagy was characterized by the accumulation of autophagic vacuoles, an increase in the expression of Beclin-1 and the stimulation of autophagic flux. We established that macroautophagy was sterol- and Beclin-1-dependent and was associated with cell survival rather than with cytotoxicity, as blockage of macroautophagy sensitized cells to AEBS ligands. These results show that the accumulation of sterols by AEBS ligands in MCF-7 cells induces apoptosis and macroautophagy. Collectively, these data support a therapeutic potential for selective AEBS ligands in breast cancer management and shows a mechanism that explains the induction of autophagy in MCF-7 cells by tamoxifen and other selective estrogen receptor modulators.
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Abbreviations
- AEBS:
-
antiestrogen-binding site
- AIF:
-
apopotosis-inducing factor
- Tx:
-
tamoxifen
- 4OHTx:
-
4-hydroxytamoxifen
- PBPE:
-
N-pyrrolidino-(phenylmethyphenoxy)-ethanamine,HCl
- DPPE:
-
N,N′-diethylamino-(phenylmethylphenoxy-ethanamine,HCl
- CHX:
-
cycloheximide
- Act D:
-
actinomycin D
- vit E:
-
vitamin E
- NAC:
-
N-acetylcysteine
- BHT:
-
butylated hydroxytoluene
- zymostenol:
-
5α-cholest-8-en-3βol
- desmosterol:
-
5α-cholest-5,24-dien-3βol
- zymosterol:
-
5α-cholest-8,24-dien-3βol
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Acknowledgements
This work was supported by an internal grant from INSERM, the Institut Claudius Regaud, the Conseil Regional Midi-Pyrénnées, the Institut National du Cancer through the ResisTH network and the company Affichem.
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de Medina, P., Payré, B., Boubekeur, N. et al. Ligands of the antiestrogen-binding site induce active cell death and autophagy in human breast cancer cells through the modulation of cholesterol metabolism. Cell Death Differ 16, 1372–1384 (2009). https://doi.org/10.1038/cdd.2009.62
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DOI: https://doi.org/10.1038/cdd.2009.62
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