Abstract
Resveratrol, a polyphenol found in grapes and other fruit and vegetables, is a powerful chemopreventive and chemotherapeutic molecule potentially of interest for the treatment of breast cancer. The human breast cancer cell line MCF-7, which is devoid of caspase-3 activity, is refractory to apoptotic cell death after incubation with resveratrol. Here we show that resveratrol arrests cell proliferation, triggers death and decreases the number of colonies of cells that are sensitive to caspase-3-dependent apoptosis (MCF-7casp-3) and also those that are unresponsive to it (MCF-7vc). We demonstrate that resveratrol (i) acts via multiple pathways to trigger cell death, (ii) induces caspase-dependent and caspase-independent cell death in MCF-7casp-3 cells, (iii) induces only caspase-independent cell death in MCF-7vc cells and (iv) stimulates macroautophagy. Using BECN1 and hVPS34 (human vacuolar protein sorting 34) small interfering RNAs, we demonstrate that resveratrol activates Beclin 1-independent autophagy in both cell lines, whereas cell death via this uncommon form of autophagy occurs only in MCF-7vc cells. We also show that this variant form of autophagic cell death is blocked by the expression of caspase-3, but not by its enzymatic activity. In conclusion, this study reveals that non-canonical autophagy induced by resveratrol can act as a caspase-independent cell death mechanism in breast cancer cells.
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Abbreviations
- Res:
-
resveratrol
- ATG:
-
autophagy-related genes
- C2-Cer:
-
C2-ceramide
- GFP:
-
green fluorescent protein
- LC3:
-
light chain 3
- 3-MA:
-
3-methyladenine
- Vps34:
-
vacuolar protein sorting 34
- PBS:
-
phosphate-buffered saline
- siRNA:
-
small interfering RNA
- PIC:
-
protease inhibitor mixture
- PARP:
-
poly(ADP-ribose) polymerase
- PI:
-
propidium iodide
- Pep A:
-
pepstatin A
- Ac-DEVD-pNA:
-
acetyl-Asp-Glu-Val-Asp p-nitroanilide
- PKB:
-
protein kinase B
- PI3K:
-
phosphoinositide 3-kinase
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Acknowledgements
We thank T Yoshimori for providing us GFP-LC3, PT Daniel for sharing human transfectant MCF-7vc and MCF-7casp-3 cells and William Dunn (University of Florida, Gainesville, FL, USA) for the Atg7 antibody. We are especially indebted to Riccarda Granata (University of Turin, Italy) for inestimable suggestions and critical reading of the manuscript. We wish to thank Dr. Iacopo Gesmundo (University of Turin) for excellent technical assistance with western blot. This study was supported by fellowships from FIRC (to Francesca Scarlatti) and University of Milan (to Roberta Maffei), by Institutional grants from University of Milan, INSERM and Université Paris Sud 11, and by research grants from the Association pour la Recherche sur le Cancer (no. 3503) in P Codogno's laboratory and from Fondazione Cariplo in R Ghidoni's laboratory.
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While this manuscript was being revised, sirtuin 1, the target of resveratrol, was shown to regulate macroautophagy (Lee et al. Proc Natl Acad Sci USA 2008; 105: 3374–3379).
Supplementary Information accompanies the paper on Cell Death and Differentiation website (http://www.nature.com/cdd)
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Scarlatti, F., Maffei, R., Beau, I. et al. Role of non-canonical Beclin 1-independent autophagy in cell death induced by resveratrol in human breast cancer cells. Cell Death Differ 15, 1318–1329 (2008). https://doi.org/10.1038/cdd.2008.51
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DOI: https://doi.org/10.1038/cdd.2008.51
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