Breast Cancer Research and Treatment

, Volume 146, Issue 3, pp 525–534 | Cite as

Metabolic differences in breast cancer stem cells and differentiated progeny

  • Erina Vlashi
  • Chann Lagadec
  • Laurent Vergnes
  • Karen Reue
  • Patricia Frohnen
  • Mabel Chan
  • Yazeed Alhiyari
  • Milana Bochkur Dratver
  • Frank PajonkEmail author
Preclinical study


In general, tumor cells display a more glycolytic phenotype compared to the corresponding normal tissue. However, it is becoming increasingly clear that tumors are composed of a heterogeneous population of cells. Breast cancers are organized in a hierarchical manner, with the breast cancer stem cells (BCSCs) at the top of the hierarchy. Here, we investigate the metabolic phenotype of BCSCs and their differentiated progeny. In addition, we determine the effect of radiation on the metabolic state of these two cell populations. Luminal, basal, and claudin-low breast cancer cell lines were propagated as mammospheres enriched in BCSCs. Lactate production, glucose consumption, and ATP content were compared with differentiated cultures. A metabolic flux analyzer was used to determine the oxygen consumption, extracellular acidification rates, maximal mitochondria capacity, and mitochondrial proton leak. The effect of radiation treatment of the metabolic phenotype of each cell population was also determined. BCSCs consume more glucose, produce less lactate, and have higher ATP content compared to their differentiated progeny. BCSCs have higher maximum mitochondrial capacity and mitochondrial proton leak compared to their differentiated progeny. Radiation treatment enhances the higher energetic state of the BCSCs, while decreasing mitochondrial proton leak. Our study indicated that breast cancer cells are heterogeneous in their metabolic phenotypes and BCSCs reside in a distinct metabolic state compared to their differentiated progeny. BCSCs display a reliance on oxidative phosphorylation, while the more differentiated progeny displays a more glycolytic phenotype. Radiation treatment affects the metabolic state of BCSCs. We conclude that interfering with the metabolic requirements of BCSCs may prevent radiation-induced reprogramming of breast cancer cells during radiation therapy, thus improving treatment outcome.


Cancer stem cells Metabolism Radiation 



We would like to thank Ekaterini Angelis, PhD, for careful editing of this manuscript. FP was supported by a generous gift from Steve and Cathy Fink and by grants from the National Cancer Institute (1RO1CA137110, 1R01CA161294) and the Army Medical Research & Materiel Command’s Breast Cancer Research Program (W81XWH-11-1-0531). LV and KR were supported by S10RR026744 (National Center for Research Resources) and P01 HL028481 (National Institutes of Health).

Conflict of interest

The authors have no conflicts of interest to disclose.

Supplementary material

10549_2014_3051_MOESM1_ESM.tiff (507 kb)
Fig. S1 a Representative oxygen consumption rates measured as a function of time on a Seahorse platform, as different metabolic inhibitors are added to the cell media. b Several parameters were deducted from the changes in oxygen consumption (a), such as: basal OCR, maximum mitochondrial capacity, and mitochondrial reserve capacity (=[maximum mitochondrial capacity] − [basal OCR]) as described previously in [10]. BCSCs and non-tumorigenic cells did not differ in ATP turnover, mitochondrial reserve capacity, or non-mitochondrial respiration


  1. 1.
    Overgaard M, Jensen MB, Overgaard J, Hansen PS, Rose C, Andersson M, Kamby C, Kjaer M, Gadeberg CC, Rasmussen BB et al (1999) Postoperative radiotherapy in high-risk postmenopausal breast-cancer patients given adjuvant tamoxifen: Danish Breast Cancer Cooperative Group DBCG 82c randomised trial. Lancet 353(9165):1641–1648PubMedCrossRefGoogle Scholar
  2. 2.
    Clarke M, Collins R, Darby S, Davies C, Elphinstone P, Evans E, Godwin J, Gray R, Hicks C, James S et al (2005) Effects of radiotherapy and of differences in the extent of surgery for early breast cancer on local recurrence and 15-year survival: an overview of the randomised trials. Lancet 366(9503):2087–2106PubMedCrossRefGoogle Scholar
  3. 3.
    Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF (2003) Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA 100:3983–3988PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Reya T, Morrison SJ, Clarke MF, Weissman IL (2001) Stem cells, cancer, and cancer stem cells. Nature 414(6859):105–111PubMedCrossRefGoogle Scholar
  5. 5.
    Warburg O, Posener K, Negelein E (1924) Ueber den Stoffwechsel der Tumoren. Biochem Z 152:319–344Google Scholar
  6. 6.
    Ter-Pogossian MM, Phelps ME, Hoffman EJ, Mullani NA (1975) A positron-emission transaxial tomograph for nuclear imaging (PETT). Radiology 114(1):89–98PubMedCrossRefGoogle Scholar
  7. 7.
    Lagadec C, Vlashi E, Della Donna L, Meng Y, Dekmezian C, Kim K, Pajonk F (2010) Survival and self-renewing capacity of breast cancer initiating cells during fractionated radiation treatment. Breast cancer Res 12(1):R13PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Vlashi E, Kim K, Lagadec C, Donna LD, McDonald JT, Eghbali M, Sayre JW, Stefani E, McBride W, Pajonk F (2009) In vivo imaging, tracking, and targeting of cancer stem cells. J Natl Cancer Inst 101(5):350–359PubMedCentralPubMedCrossRefGoogle Scholar
  9. 9.
    Pollard SM, Yoshikawa K, Clarke ID, Danovi D, Stricker S, Russell R, Bayani J, Head R, Lee M, Bernstein M et al (2009) Glioma stem cell lines expanded in adherent culture have tumor-specific phenotypes and are suitable for chemical and genetic screens. Cell Stem Cell 4(6):568–580PubMedCrossRefGoogle Scholar
  10. 10.
    Amo T, Yadava N, Oh R, Nicholls DG, Brand MD (2008) Experimental assessment of bioenergetic differences caused by the common European mitochondrial DNA haplogroups H and T. Gene 411(1–2):69–76PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Lagadec C, Vlashi E, Della Donna L, Dekmezian C, Pajonk F (2012) Radiation-induced reprogramming of breast cancer cells. Stem cells 30(5):833–844PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Lagadec C, Vlashi E, Alhiyari Y, Phillips TM, Dratver MB, Pajonk F (2013) Radiation-induced notch signaling in breast cancer stem cells. Int J Radiat Oncol Biol Phys 87(3):609PubMedCrossRefGoogle Scholar
  13. 13.
    Vlashi E, Lagadec C, Chan M, Frohnen P, McDonald AJ, Pajonk F (2013) Targeted elimination of breast cancer cells with low proteasome activity is sufficient for tumor regression. Breast Cancer Res Treat 141(2):197–203PubMedCrossRefGoogle Scholar
  14. 14.
    Conley SJ, Gheordunescu E, Kakarala P, Newman B, Korkaya H, Heath AN, Clouthier SG, Wicha MS (2012) Antiangiogenic agents increase breast cancer stem cells via the generation of tumor hypoxia. Proc Natl Acad Sci USA 109(8):2784–2789PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA, Daidone MG (2005) Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res 65(13):5506–5511PubMedCrossRefGoogle Scholar
  16. 16.
    Christofk HR, Vander Heiden MG, Harris MH, Ramanathan A, Gerszten RE, Wei R, Fleming MD, Schreiber SL, Cantley LC (2008) The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature 452(7184):230–233PubMedCrossRefGoogle Scholar
  17. 17.
    Christofk HR, Vander Heiden MG, Asara JM, Cantley LC (2008) Pyruvate kinase M2 is a phosphotyrosine-binding protein. Nature 452(7184):181–186PubMedCrossRefGoogle Scholar
  18. 18.
    Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676PubMedCrossRefGoogle Scholar
  19. 19.
    Moon JS, Kim HE, Koh E, Park SH, Jin WJ, Park BW, Park SW, Kim KS (2011) Kruppel-like factor 4 (KLF4) activates the transcription of the gene for the platelet isoform of phosphofructokinase (PFKP) in breast cancer. J Biol Chem 286(27):23808–23816PubMedCentralPubMedCrossRefGoogle Scholar
  20. 20.
    Vlashi E, Lagadec C, Vergnes L, Matsutani T, Masui K, Poulou M, Della Donna L, Evers P et al (2011) Metabolic state of glioma stem cells and nontumorigenic cells. Proc Natl Acad Sci USA 108(38):16062–16067PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Warburg O (1924) On the metabolism of carcinoma cells. Biochem Z 152(309–344):309Google Scholar
  22. 22.
    Lagadinou ED, Sach A, Callahan K, Rossi RM, Neering SJ, Minhajuddin M, Ashton JM, Pei S, Grose V, O’Dwyer KM et al (2013) BCL-2 inhibition targets oxidative phosphorylation and selectively eradicates quiescent human leukemia stem cells. Cell Stem Cell 12(3):329–341PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Vander Heiden MG, Cantley LC, Thompson CB (2009) Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324(5930):1029–1033PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Zhang WC, Shyh-Chang N, Yang H, Rai A, Umashankar S, Ma S, Soh BS, Sun LL, Tai BC, Nga ME et al (2012) Glycine decarboxylase activity drives non-small cell lung cancer tumor-initiating cells and tumorigenesis. Cell 148(1–2):259–272PubMedCrossRefGoogle Scholar
  25. 25.
    Morfouace M, Lalier L, Bahut M, Bonnamain V, Naveilhan P, Guette C, Oliver L, Gueguen N, Reynier P, Vallette FM (2012) Comparison of spheroids formed by rat glioma stem cells and neural stem cells reveals differences in glucose metabolism and promising therapeutic applications. J Biol Chem 287(40):33664–33674PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Menendez JA, Joven J, Cufi S, Corominas-Faja B, Oliveras-Ferraros C, Cuyas E, Martin-Castillo B, Lopez-Bonet E, Alarcon T, Vazquez-Martin A (2013) The Warburg effect version 2.0: metabolic reprogramming of cancer stem cells. Cell Cycle 12(8):1166–1179PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Feng W, Gentles A, Nair RV, Huang M, Lin Y, Lee CY, Cai S, Scheeren FA, Kuo AH, Diehn M (2014) Targeting unique metabolic properties of breast tumor initiating cells. Stem cells 32(7):1734–1745PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Erina Vlashi
    • 1
    • 2
  • Chann Lagadec
    • 1
  • Laurent Vergnes
    • 3
  • Karen Reue
    • 3
  • Patricia Frohnen
    • 1
  • Mabel Chan
    • 1
  • Yazeed Alhiyari
    • 1
  • Milana Bochkur Dratver
    • 1
  • Frank Pajonk
    • 1
    • 2
    Email author
  1. 1.Department of Radiation OncologyDavid Geffen School of Medicine at UCLALos AngelesUSA
  2. 2.Jonsson Comprehensive Cancer Center at UCLALos AngelesUSA
  3. 3.Department of Human GeneticsDavid Geffen School of Medicine at UCLALos AngelesUSA

Personalised recommendations