Summary
Apoptosis was induced by treating L1210 leukaemia cells with mechlorethamine, and SW620 colorectal cells with doxorubicin. The onset and progression of apoptosis were monitored by assessing caspase activation, mitochondrial transmembrane potential, phosphatidylserine externalization, DNA fragmentation and cell morphology. In parallel, 31P magnetic resonance (MR) spectra of cell extracts were recorded. In L1210 cells, caspase activation was detected at 4 h. By 3 h, the MR spectra showed a steady decrease in NTP and NAD, and a significant build-up of fructose 1,6-bisphosphate (F-1,6-P) dihydroxyacetonephosphate and glycerol-3-phosphate, indicating modulation of glycolysis. Treatment with iodoacetate also induced a build-up of F-1,6-P, while preincubation with two poly(ADP-ribose) polymerase inhibitors, 3-aminobenzamide and nicotinamide, prevented the drop in NAD and the build-up of glycolytic intermediates. This suggested that our results were due to inhibition of glyceraldehyde-3-phosphate dehydrogenase, possibly as a consequence of NAD depletion following poly(ADP-ribose) polymerase activation. Doxorubicin treatment of the adherent SW620 cells caused cells committed to apoptosis to detach. F-1,6-P was observed in detached cells, but not in treated cells that remained attached. This indicated that our observations were not cell line- or treatment-specific, but were correlated with the appearance of apoptotic cells following drug treatment. The 31P MR spectrum of tumours responding to chemotherapy could be modulated by similar effects.
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Adebodun, F. & Post, J. F. (1994). 31P NMR characterization of cellular metabolism during dexamethasone induced apoptosis in human leukemic cell lines. J Cell Physiol 158: 180–186.
Berners-Price-SJ,, Sant, M. E., Christopherson, R. I. & Kuchel, P. W. (1991). 1H and 31P NMR and HPLC studies of mouse L1210 leukemia cell extracts: the effect of Au(I) and Cu(I) diphosphine complexes on the cell metabolism. Magn Reson Med 18: 142–158.
Eguchi, Y., Shimizu, S. & Tsujimoto, Y. (1997). Intracellular ATP levels determine cell death fate by apoptosis or necrosis. Cancer Res 57: 1835–1840.
Eigenbrodt, E., Gerbracht, U., Mazurek, S., Presket, P. & Friis, R. (1994). Carbohydrate metabolism and neoplasia: new perspectives for diagnosis and therapy. In Biochemical and Molecular Aspects of Selected Cancers, Prestlow TG and Prestlow TP(eds), pp. 311–385. Academic Press, San Diego
Ellis, P. A., Smith, I. E., McCarthy, K., Detre, S., Salter, J. & Dowsett, M. (1997). Preoperative chemotherapy induces apoptosis in early breast cancer [letter]. Lancet 349: 849
Hague, A., Manning, A. M., Hanlon, K. A., Huschtscha, L. I., Hart, D. & Paraskeva, C. (1993). Sodium butyrate induces apoptosis in human colonic tumour cell lines in a p-53 independent pathway: implications for the possible role of dietary fibre in the prevention of large-bowel cancer. Int J Cancer 55: 498–505.
Heerdt, B. G., Houston, M. A. & Augenlich, L. H. (1994). Potentiation by specific short-chain fatty acids of differentiation and apoptosis in human colonic carcinoma cell lines. Cancer Res 54: 3288–3294.
Hickman, J. A. (1992). Apoptosis induced by anticancer drugs. Cancer Metastasis Rev 11: 121–139.
Hill, M. E., MacLennan, K. A., Cunningham, D. C., Vaughan-Hudson, B., Burke, M., Clarke, P., Di-Stefano, F., Anderson, L., Vaughan-Hudson, G., Mason, D., Selby, P. & Linch, D. C. (1996). Prognostic significance of bcl-2 expression and bcl-2 major breakpoint region rearrangement in diffuse large cell non-Hodgkin’s lymphoma: a British National Lymphoma Investigation Study. Blood 88: 1046–1051.
Koch-Nolte, F. & Haag, F. (1997). Mono(ADP-ribosyl)transferases and related enzymes in animal tissues. Emerging gene families. Adv Exp Med Biol 419: 1–13.
Leist, M., Single, B., Kunstle, G., Volbracht, C., Hentze, H. & Nicotera, P. (1997). Apoptosis in the absence of poly-(ADP-ribose) polymerase. Biochem Biophys Res Commun 233: 518–522.
Martin, D. S., Stolfi, R. L. & Colofiore, J. R. (1997). Perspective: the chemotherapeutic relevance of apoptosis and a proposed biochemical cascade for chemotherapeutically induced apoptosis. Cancer Invest 15: 372–381.
Milam, K. M. & Cleaver, J. E. (1984). Inhibitors of poly(adenosine diphosphate-ribose) synthesis: effect on other metabolic processes. Science 223: 589–591.
Mueller, S. G., Schraw, W. P. & Richmond, A. (1995). Activation of protein kinase C enhances the phosphorylation of the type B interleukin-8 receptor and stimulates its degradation in non-hematopoietic cells. J Biol Chem 270: 10439–10448.
Neeman, M., Eldar, H., Rushkin, E. & Degani, H. (1990). Chemotherapy-induced changes in the energetics of human breast cancer cells; 31P- and 13C-NMR studies. Biochim Biophys Acta 1052: 255–263.
Negendank, W., Padavic Shaller, K., Li, C. W., Murphy Boesch, J., Schilder, R. S., Smith, M. R. & Brown, T. R. (1996). Phosphomonoester changes in vivo and treatment response in human non-Hodgkin’s lymphomas (meeting abstract). Proc Annu Meet Am Assoc Cancer Res 37: A1343
Nosseri, C., Coppola, S. & Ghibelli, L. (1994). Possible involvement of poly(ADP-ribosyl) polymerase in triggering stress-induced apoptosis. Exp Cell Res 212: 367–373.
Nunn, A. V., Barnard, M. L., Bhakoo, K., Murray, J., Chilvers, E. J. & Bell, J. D. (1996). Characterisation of secondary metabolites associated with neutrophil apoptosis. FEBS Lett 392: 295–298.
O’Connor, P. M., Jackman, J., Bae, I., Myers, T. G., Fan, S., Mutoh, M., Scudiero, D. A., Monks, A., Sausville, E. A., Weinstein, J. N., Friend, S., Fornace, A. J. Jr & Kohn, K. W. (1997). Characterization of the p53 tumor suppressor pathway in cell lines of the National Cancer Institute anticancer drug screen and correlations with the growth-inhibitory potency of 123 anticancer agents. Cancer Res 57: 4285–4300.
Ronen, S. M., Stier, A. & Degani, H. (1990). NMR studies of the lipid metabolism of T47D human breast cancer spheroids. FEBS 266: 147–150.
Satoh, M. S. & Lindahl, T. (1992). Role of poly(ADP-ribose) formation in DNA repair. Nature 356: 356–358.
Tyagi, R. K., Azrad, A., Degani, H. & Salomon, Y. (1996). Simultaneous extraction of cellular lipids and water-soluble metabolites: evaluation by NMR spectroscopy. Magn Reson Med 35: 194–200.
Webb, J. L. (1966). Iodoacetate and iodoacetamide. In: Enzyme and Metabolic Inhibitors, pp. 1–270. Academic Press: New York
White, A., Handler, P., Smith, E. L., Hill, R. L. & Lehman, I. R. (1978). Principles of Biochemistry, 6th edn. McGraw-Hill Kogakusha: Tokyo
White, E. (1996). Life, death, and the pursuit of apoptosis. Genes Dev 10: 1–15.
Zamzami, N., Marchetti, P., Castedo, M., Zanin, C., Vayssiere, J-L, Petit, P. X. & Kroemer, G. (1995). Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo. J Exp Med 181: 1661–1672.
Zhang, J., Dawson, T. M., Dawson, V. L. & Snyder, S. H. (1994). Nitric oxide activation of poly(ADP-ribose) synthetase in neurotoxicity. Science 263: 687–689.
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Ronen, S., DiStefano, F., McCoy, C. et al. Magnetic resonance detects metabolic changes associated with chemotherapy-induced apoptosis. Br J Cancer 80, 1035–1041 (1999). https://doi.org/10.1038/sj.bjc.6690459
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.bjc.6690459
Keywords
This article is cited by
-
In vivo magnetic resonance imaging of treatment-induced apoptosis
Scientific Reports (2019)
-
17-allyamino-17-demethoxygeldanamycin treatment results in a magnetic resonance spectroscopy-detectable elevation in choline-containing metabolites associated with increased expression of choline transporter SLC44A1 and phospholipase A2
Breast Cancer Research (2010)
-
From metabolic to metabolomic NMR spectroscopy of apoptotic cells
Metabolomics (2005)
-
Apoptosis is associated with triacylglycerol accumulation in Jurkat T-cells
British Journal of Cancer (2002)
-
Early gene therapy–induced apoptotic response in BT4C gliomas by magnetic resonance relaxation contrast T1 in the rotating frame
Cancer Gene Therapy (2002)