Normal epithelial cells require matrix attachment for survival, and the ability of tumour cells to survive outside their natural extracellular matrix (ECM) niches is dependent on acquisition of anchorage independence1. Although apoptosis is the most rapid mechanism for eliminating cells lacking appropriate ECM attachment2, recent reports suggest that non-apoptotic death processes prevent survival when apoptosis is inhibited in matrix-deprived cells3,4. Here we demonstrate that detachment of mammary epithelial cells from ECM causes an ATP deficiency owing to the loss of glucose transport. Overexpression of ERBB2 rescues the ATP deficiency by restoring glucose uptake through stabilization of EGFR and phosphatidylinositol-3-OH kinase (PI(3)K) activation, and this rescue is dependent on glucose-stimulated flux through the antioxidant-generating pentose phosphate pathway. Notably, we found that the ATP deficiency could be rescued by antioxidant treatment without rescue of glucose uptake. This rescue was found to be dependent on stimulation of fatty acid oxidation, which is inhibited by detachment-induced reactive oxygen species (ROS). The significance of these findings was supported by evidence of an increase in ROS in matrix-deprived cells in the luminal space of mammary acini, and the discovery that antioxidants facilitate the survival of these cells and enhance anchorage-independent colony formation. These results show both the importance of matrix attachment in regulating metabolic activity and an unanticipated mechanism for cell survival in altered matrix environments by antioxidant restoration of ATP generation.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Simpson, C. D., Anyiwe, K. & Schimmer, A. D. Anoikis resistance and tumor metastasis. Cancer Lett. 272, 177–185 (2008)
Frisch, S. M. & Screaton, R. A. Anoikis mechanisms. Curr. Opin. Cell Biol. 13, 555–562 (2001)
Debnath, J. et al. The role of apoptosis in creating and maintaining luminal space within normal and oncogene-expressing mammary acini. Cell 111, 29–40 (2002)
Mailleux, A. A. et al. BIM regulates apoptosis during mammary ductal morphogenesis, and its absence reveals alternative cell death mechanisms. Dev. Cell 12, 221–234 (2007)
Nelson, C. M. & Bissell, M. J. Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer. Annu. Rev. Cell Dev. Biol. 22, 287–309 (2006)
Debnath, J. & Brugge, J. S. Modelling glandular epithelial cancers in three-dimensional cultures. Nature Rev. Cancer 5, 675–688 (2005)
Humphreys, R. C. et al. Apoptosis in the terminal endbud of the murine mammary gland: a mechanism of ductal morphogenesis. Development 122, 4013–4022 (1996)
Muthuswamy, S. K. et al. ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini. Nature Cell Biol. 3, 785–792 (2001)
Fung, C. et al. Induction of autophagy during extracellular matrix detachment promotes cell survival. Mol. Biol. Cell 19, 797–806 (2008)
Mills, K. R. et al. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is required for induction of autophagy during lumen formation in vitro. Proc. Natl Acad. Sci. USA 101, 3438–3443 (2004)
Mathew, R., Karantza-Wadsworth, V. & White, E. Role of autophagy in cancer. Nature Rev. Cancer 7, 961–967 (2007)
Reginato, M. J. et al. Integrins and EGFR coordinately regulate the pro-apoptotic protein Bim to prevent anoikis. Nature Cell Biol. 5, 733–740 (2003)
Czech, M. P. & Corvera, S. Signaling mechanisms that regulate glucose transport. J. Biol. Chem. 274, 1865–1868 (1999)
Elstrom, R. L. et al. Akt stimulates aerobic glycolysis in cancer cells. Cancer Res. 64, 3892–3899 (2004)
DeBerardinis, R. J., Lum, J. J., Hatzivassiliou, G. & Thompson, C. B. The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. Cell Metab. 7, 11–20 (2008)
Bensaad, K. et al. TIGAR, a p53-inducible regulator of glycolysis and apoptosis. Cell 126, 107–120 (2006)
Boada, J. et al. Cells overexpressing fructose-2,6-bisphosphatase showed enhanced pentose phosphate pathway flux and resistance to oxidative stress. FEBS Lett. 480, 261–264 (2000)
Li, A. E. et al. A role for reactive oxygen species in endothelial cell anoikis. Circ. Res. 85, 304–310 (1999)
Buzzai, M. et al. The glucose dependence of Akt-transformed cells can be reversed by pharmacologic activation of fatty acid β-oxidation. Oncogene 24, 4165–4173 (2005)
Deberardinis, R. J., Lum, J. J. & Thompson, C. B. Phosphatidylinositol 3-kinase-dependent modulation of carnitine palmitoyltransferase 1A expression regulates lipid metabolism during hematopoietic cell growth. J. Biol. Chem. 281, 37372–37380 (2006)
Bennett, B. D. et al. Quantitative subcellular imaging of glucose metabolism within intact pancreatic islets. J. Biol. Chem. 271, 3647–3651 (1996)
Chance, B., Cohen, P., Jobsis, F. & Schoener, B. Intracellular oxidation-reduction states in vivo. Science 137, 499–508 (1962)
Gao, P. et al. HIF-dependent antitumorigenic effect of antioxidants in vivo. Cancer Cell 12, 230–238 (2007)
Narayanan, B. A. Chemopreventive agents alters global gene expression pattern: predicting their mode of action and targets. Curr. Cancer Drug Targets 6, 711–727 (2006)
Ivshina, A. V. et al. Genetic reclassification of histologic grade delineates new clinical subtypes of breast cancer. Cancer Res. 66, 10292–10301 (2006)
Sorlie, T. et al. Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications. Proc. Natl Acad. Sci. USA 98, 10869–10874 (2001)
Chen, E. I. et al. Adaptation of energy metabolism in breast cancer brain metastases. Cancer Res. 67, 1472–1486 (2007)
Omenn, G. S. et al. Effects of a combination of β carotene and vitamin A on lung cancer and cardiovascular disease. N. Engl. J. Med. 334, 1150–1155 (1996)
Lawenda, B. D. et al. Should supplemental antioxidant administration be avoided during chemotherapy and radiation therapy? J. Natl Cancer Inst. 100, 773–783 (2008)
Debnath, J., Muthuswamy, S. K. & Brugge, J. S. Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods 30, 256–268 (2003)
Overholtzer, M. et al. Transforming properties of YAP, a candidate oncogene on the chromosome 11q22 amplicon. Proc. Natl Acad. Sci. USA 103, 12405–12410 (2006)
Schafer, Z. T. et al. Enhanced sensitivity to cytochrome c-induced apoptosis mediated by PHAPI in breast cancer cells. Cancer Res. 66, 2210–2218 (2006)
Sebastia, J. et al. Evaluation of fluorescent dyes for measuring intracellular glutathione content in primary cultures of human neurons and neuroblastoma SH-SY5Y. Cytometry A 51, 16–25 (2003)
Vishwasrao, H. D., Heikal, A. A., Kasischke, K. A. & Webb, W. W. Conformational dependence of intracellular NADH on metabolic state revealed by associated fluorescence anisotropy. J. Biol. Chem. 280, 25119–25126 (2005)
We thank M. Overholtzer, G. Mouneimne, M. Mazzone and C. Leung for critical reading of the manuscript and/or experimental assistance. We thank A. Mailleux, A. Kaanta, V. Schafer, A. Zhou, K. Simpson, and the members of the Brugge laboratory for experimental assistance, comments, and/or discussion. This work was supported by a grant from the National Cancer Institute (J.S.B.) and a grant from the National Institutes of Health (NIH) (P.P.). Z.T.S. is the recipient of a Ruth L. Kirschstein National Research Service Award (NRSA) for Postdoctoral Fellows from the National Cancer Institute; L.S. an NCI Mentored Quantitative Research Development Award (K25); A.R.G. a National Science Graduate Research Fellowship; and H.Y.I. an NCI K08 Award.
Author Contributions Z.T.S. and J.S.B. were responsible for the overall study design. Z.T.S., A.R.G., H.Y.I. and S.G. conducted experiments. L.S. and Z.J. conducted the experiments measuring native fluorescence of NADPH in 3D cell culture. Z.G.-H. and P.P. designed the fatty acid oxidation studies and Z.T.S. and Z.G.-H. conducted the fatty acid oxidation assays. Z.T.S. and J.S.B. drafted the manuscript and all other authors made revisions.
About this article
Cite this article
Schafer, Z., Grassian, A., Song, L. et al. Antioxidant and oncogene rescue of metabolic defects caused by loss of matrix attachment. Nature 461, 109–113 (2009) doi:10.1038/nature08268
Future Medicinal Chemistry (2019)
Precision Clinical Medicine (2019)
European Journal of Medicinal Chemistry (2019)
3D collagen architecture regulates cell adhesion through degradability, thereby controlling metabolic and oxidative stress
Integrative Biology (2019)