Abstract
We recently reported that shikonin and its analogs were a class of necroptotic inducers that could bypass cancer drug resistance. However, the molecular targets of shikonin are not known. Here, we showed that shikonin and its analogs are inhibitors of tumor-specific pyruvate kinase-M2 (PKM2), among which shikonin and its enantiomeric isomer alkannin were the most potent and showed promising selectivity, that is, shikonin and alkannin at concentrations that resulted in over 50% inhibition of PKM2 activity did not inhibit PKM1 and pyruvate kinase-L (PKL). Shikonin and alkannin significantly inhibited the glycolytic rate, as manifested by cellular lactate production and glucose consumption in drug-sensitive and resistant cancer cell lines (MCF-7, MCF-7/Adr, MCF-7/Bcl-2, MCF-7/Bcl-xL and A549) that primarily express PKM2. HeLa cells transfected with PKM1 showed reduced sensitivity to shikonin- or alkannin-induced cell death. To the best of our knowledge, shikonin and alkannin are the most potent and specific inhibitors to PKM2 reported so far. As PKM2 universally expresses in cancer cells and dictates the last rate-limiting step of glycolysis vital for cancer cell proliferation and survival, enantiomeric shikonin and alkannin may have potential in future clinical application.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
Ahn BZ, Baik KU, Kweon GR, Lim K, Hwang BD . (1995). Acylshikonin analogues: synthesis and inhibition of DNA topoisomerase-I. J Med Chem 38: 1044–1047.
Altenberg B, Greulich KO . (2004). Genes of glycolysis are ubiquitously overexpressed in 24 cancer classes. Genomics 84: 1014–1020.
Anderson SR, Florini JR, Vestling CS . (1964). Rat liver lactate dehydrogenase. 3. Kinetics and specificity. J Biol Chem 239: 2991–2997.
Bailly C . (2000). Topoisomerase I poisons and suppressors as anticancer drugs. Curr Med Chem 7: 20.
Christofk HR, Vander Heiden MG, Harris MH, Ramanathan A, Gerszten RE, Wei R et al. (2008). The M2 splice isoform of pyruvate kinase is important for cancer metabolism and tumour growth. Nature 452: 230–233.
DeBerardinis RJ, Lum JJ, Hatzivassiliou G, Thompson CB . (2008). The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. Cell Metab 7: 11–20.
Degterev A, Huang Z, Boyce M, Li Y, Jagtap P, Mizushima N et al. (2005). Chemical inhibitor of nonapoptotic cell death with therapeutic potential for ischemic brain injury. Nat Chem Biol 1: 112–119.
Degterev A, Hitomi J, Germscheid M, Ch′en IL, Korkina O, Teng X et al. (2008). Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol 4: 313–321.
Dombrauckas JD, Santarsiero BD, Mesecar AD . (2005). Structural basis for tumor pyruvate kinase M2 allosteric regulation and catalysis. Biochemistry 44: 9417–9429.
Gatenby RA, Gillies RJ . (2004). Why do cancers have high aerobic glycolysis? Nat Rev Cancer 4: 891–899.
Guo XP, Zhang XY, Zhang SD . (1991). [Clinical trial on the effects of shikonin mixture on later stage lung cancer]. Zhong Xi Yi Jie He Za Zhi 11: 598–599, 580.
Han W, Li L, Qiu S, Lu Q, Pan Q, Gu Y et al. (2007). Shikonin circumvents cancer drug resistance by induction of a necroptotic death. Mol Cancer Ther 6: 1641–1649.
Han W, Xie J, Li L, Liu Z, Hu X . (2009). Necrostatin-1 reverts shikonin-induced necroptosis to apoptosis. Apoptosis 14: 674–686.
Hirayama A, Kami K, Sugimoto M, Sugawara M, Toki N, Onozuka H et al. (2009). Quantitative metabolome profiling of colon and stomach cancer microenvironment by capillary electrophoresis time-of-flight mass spectrometry. Cancer Res 69: 4918–4925.
Hitomi J, Christofferson DE, Ng A, Yao J, Degterev A, Xavier RJ et al. (2008). Identification of a molecular signaling network that regulates a cellular necrotic cell death pathway. Cell 135: 1311–1323.
Hsu PP, Sabatini DM . (2008). Cancer cell metabolism: Warburg and beyond. Cell 134: 703–707.
Hu X, Xuan Y . (2008). Bypassing cancer drug resistance by activating multiple death pathways—a proposal from the study of circumventing cancer drug resistance by induction of necroptosis. Cancer Lett 259: 127–137.
Ibsen KH . (1977). Interrelationships and functions of the pyruvate kinase isozymes and their variant forms: a review. Cancer Res 37: 341–353.
Ikeda Y, Noguchi T . (1998). Allosteric regulation of pyruvate kinase M2 isozyme involves a cysteine residue in the intersubunit contact. J Biol Chem 273: 12227–12233.
Imamura K, Tanaka T, Nishina T, Nakashima K, Miwa S . (1973). Studies on pyruvate kinase (PK) deficiency. II. Electrophoretic, kinetic and immunological studies on pyruvate kinase of erythrocytes and other tissues. J Biochem 74: 1165–1175.
Jurica MS, Mesecar A, Heath PJ, Shi W, Nowak T, Stoddard BL . (1998). The allosteric regulation of pyruvate kinase by fructose-1,6-bisphosphate. Structure 6: 195–210.
Kim SH, Kang IC, Yoon TJ, Park YM, Kang KS, Song GY et al. (2001). Antitumor activities of a newly synthesized shikonin derivative, 2-hyim-DMNQ-S-33. Cancer Lett 172: 171–175.
Li L, Han W, Gu Y, Qiu S, Lu Q, Jin J et al. (2007). Honokiol induces a necrotic cell death through the mitochondrial permeability transition pore. Cancer Res 67: 4894–4903.
Liu K, Lu H, Hou L, Qi Z, Teixeira C, Barbault F et al. (2008). Design, synthesis, and biological evaluation of N-carboxyphenylpyrrole derivatives as potent HIV fusion inhibitors targeting gp41. J Med Chem 51: 7843–7854.
Masuda Y, Nishida A, Hori K, Hirabayashi T, Kajimoto S, Nakajo S et al. (2003). Beta-hydroxyisovalerylshikonin induces apoptosis in human leukemia cells by inhibiting the activity of a polo-like kinase 1 (PLK1). Oncogene 22: 1012–1023.
Masuda Y, Shima G, Aiuchi T, Horie M, Hori K, Nakajo S et al. (2004). Involvement of tumor necrosis factor receptor-associated protein 1 (TRAP1) in apoptosis induced by beta-hydroxyisovalerylshikonin. J Biol Chem 279: 42503–42515.
Mazurek S . (2007). Pyruvate kinase type M2: a key regulator within the tumour metabolome and a tool for metabolic profiling of tumours. Ernst Schering Found Symp Proc 4: 99–124.
Mazurek S, Boschek CB, Hugo F, Eigenbrodt E . (2005). Pyruvate kinase type M2 and its role in tumor growth and spreading. Semin Cancer Biol 15: 300–308.
Mazurek S, Eigenbrodt E . (2003). The tumor metabolome. Anticancer Res 23: 1149–1154.
Mazurek S, Grimm H, Boschek CB, Vaupel P, Eigenbrodt E . (2002). Pyruvate kinase type M2: a crossroad in the tumor metabolome. Br J Nutr 87 (Suppl 1): S23–S29.
Nakaya K, Miyasaka T . (2003). A shikonin derivative, beta-hydroxyisovalerylshikonin, is an ATP-non-competitive inhibitor of protein tyrosine kinases. Anticancer Drugs 14: 683–693.
Noguchi T, Inoue H, Tanaka T . (1986). The M1- and M2-type isozymes of rat pyruvate kinase are produced from the same gene by alternative RNA splicing. J Biol Chem 261: 13807–13812.
Noguchi T, Yamada K, Inoue H, Matsuda T, Tanaka T . (1987). The L- and R-type isozymes of rat pyruvate kinase are produced from a single gene by use of different promoters. J Biol Chem 262: 14366–14371.
Papageorgiou VP AA, Couladouros EA, Hepworth D, Nicolaou KC. . (1999). The chemistry and biology of alkannin, shikonin, and related naphthazarin natural products. Angewandte Chemie International Edition 38: 32.
Parkison C, Ashizawa K, McPhie P, Lin K, Cheng S . (1991). The monomer of pyruvate kinase, subtype M1, is both a kinase and a cytosolic thyroid hormone binding protein. Biochem Biophys Res Commun 179: 668–674.
Shimada N, Shinagawa T, Ishii S . (2008). Modulation of M2-type pyruvate kinase activity by the cytoplasmic PML tumor suppressor protein. Genes Cells 13: 245–254.
Spoden GA, Mazurek S, Morandell D, Bacher N, Ausserlechner MJ, Jansen-Durr P et al. (2008). Isotype-specific inhibitors of the glycolytic key regulator pyruvate kinase subtype M2 moderately decelerate tumor cell proliferation. Int J Cancer 123: 312–321.
Spoden GA, Rostek U, Lechner S, Mitterberger M, Mazurek S, Zwerschke W . (2009). Pyruvate kinase isoenzyme M2 is a glycolytic sensor differentially regulating cell proliferation, cell size and apoptotic cell death dependent on glucose supply. Exp Cell Res 315: 2765–2774.
Vander Heiden MG, Cantley LC, Thompson CB . (2009). Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324: 1029–1033.
Vander Heiden MG, Christofk HR, Schuman E, Subtelny AO, Sharfi H, Harlow EE et al. (2010). Identification of small molecule inhibitors of pyruvate kinase M2. Biochem Pharmacol 79: 1118–1124.
Warburg O . (1956). On the origin of cancer cells. Science 123: 309–314.
Xuan Y, Hu X . (2009). Naturally-occurring shikonin analogues—a class of necroptotic inducers that circumvent cancer drug resistance. Cancer Lett 274: 233–242.
Yang H, Zhou P, Huang H, Chen D, Ma N, Cui QC et al. (2009). Shikonin exerts antitumor activity via proteasome inhibition and cell death induction in vitro and in vivo. Int J Cancer 124: 2450–2459.
Acknowledgements
We thank BPS Bioscience Inc (San Diego, CA, USA) for technical support of their product—recombinant human PKM2. This work was supported in part by the China National 863 project (2007AA02Z143) to XH; China Natural Sciences Foundation projects (30772544, 81071802) to XH and the Fundamental Research Funds for the Central Universities, National Ministry of Education, China, to XH.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Supplementary information
Rights and permissions
About this article
Cite this article
Chen, J., Xie, J., Jiang, Z. et al. Shikonin and its analogs inhibit cancer cell glycolysis by targeting tumor pyruvate kinase-M2. Oncogene 30, 4297–4306 (2011). https://doi.org/10.1038/onc.2011.137
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2011.137
Keywords
This article is cited by
-
Inhibition of Pyruvate kinase M2 (PKM2) by shikonin attenuates isoproterenol-induced acute myocardial infarction via reduction in inflammation, hypoxia, apoptosis, and fibrosis
Naunyn-Schmiedeberg's Archives of Pharmacology (2024)
-
Mediation of PKM2-dependent glycolytic and non-glycolytic pathways by ENO2 in head and neck cancer development
Journal of Experimental & Clinical Cancer Research (2023)
-
Pyruvate kinase M2 regulates mitochondrial homeostasis in cisplatin-induced acute kidney injury
Cell Death & Disease (2023)
-
IgSF11-mediated phosphorylation of pyruvate kinase M2 regulates osteoclast differentiation and prevents pathological bone loss
Bone Research (2023)
-
Role of Pyruvate Kinase M2 (PKM2) in Cardiovascular Diseases
Journal of Cardiovascular Translational Research (2023)