Medical Oncology

, 36:11 | Cite as

Isocitrate dehydrogenase gene mutations and 2-hydroxyglutarate accumulation in esophageal squamous cell carcinoma

  • Keisuke Miyake
  • Yoshifumi Baba
  • Takatsugu Ishimoto
  • Yukiharu Hiyoshi
  • Masaaki Iwatsuki
  • Yuji Miyamoto
  • Naoya Yoshida
  • Masayuki Watanabe
  • Yoko Ogata
  • Megumi Nagayama
  • Atit Silsirivanit
  • Daiki Kobayashi
  • Norie Araki
  • Hideo BabaEmail author
Original Paper


Isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) are key metabolic enzymes that convert isocitrate to α-ketoglutarate. Somatic point mutations in IDH1/2 confer a gain-of-function in cancer cells, resulting in overproduction of an oncometabolite, 2-hydroxyglutarate (2HG). 2HG interferes with cellular metabolism and epigenetic regulation, contributing to oncogenesis. Given that IDH1 and IDH2 are attracting attention as promising therapeutic targets, better evaluation of the incidence of IDH1 and IDH2 mutations and 2HG level in human cancers is clinically important. This is the first study to assess their incidence in esophageal squamous cell carcinomas (ESCCs). First, we established pyrosequencing assays for IDH1 and IDH2 mutations and revealed that these mutations were absent in 10 ESCC cell lines and 96 ESCC tissues. Second, utilizing IDH1 and IDH2 overexpression vectors, we demonstrated that LC-MS/MS assays can accurately evaluate 2HG level and found that some ESCC cases presented a high level of 2HG. In conclusion, IDH1 or IDH2 mutations play a limited role in the development of ESCC. 2HG is potentially synthesized to high levels in the absence of IDH1 and IDH2 mutations, and this may correlate with progression of ESCCs.


Esophageal squamous cell carcinoma Isocitrate dehydrogenase 2-Hydroxyglutarate α-Ketoglutaric acid Oncometabolite 



Isocitrate dehydrogenase (IDH)







We thank Yuko Taniguchi and Naomi Yokoyama for technical assistance. This work was supported in part by a Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research, Grant Number 17H04273.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest.

Supplementary material

12032_2018_1229_MOESM1_ESM.docx (20 kb)
Supplementary material 1 (DOCX 19 KB)
12032_2018_1229_MOESM2_ESM.docx (21 kb)
Supplementary material 2 (DOCX 21 KB)


  1. 1.
    Lehnert T. Multimodal therapy for squamous carcinoma of the oesophagus. Br J Surg. 1999;86(6):727–39.CrossRefGoogle Scholar
  2. 2.
    Katlic MR, Wilkins EW Jr, Grillo HC. Three decades of treatment of esophageal squamous carcinoma at the Massachusetts General Hospital. J Thorac Cardiovasc Surg. 1990;99(5):929–38.PubMedGoogle Scholar
  3. 3.
    Flamen P, Van Cutsem E, Lerut A, et al. Positron emission tomography for assessment of the response to induction radiochemotherapy in locally advanced oesophageal cancer. Ann Oncol. 2002;13(3):361–8.CrossRefGoogle Scholar
  4. 4.
    Brucher BL, Weber W, Bauer M, et al. Neoadjuvant therapy of esophageal squamous cell carcinoma: response evaluation by positron emission tomography. Ann Surg. 2001;233(3):300–9.CrossRefGoogle Scholar
  5. 5.
    Downey RJ, Akhurst T, Ilson D, et al. Whole body 18FDG-PET and the response of esophageal cancer to induction therapy: results of a prospective trial. J Clin Oncol. 2003;21(3):428–32.CrossRefGoogle Scholar
  6. 6.
    Dang L, White DW, Gross S, et al. Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature. 2009;462(7274):739–44.CrossRefGoogle Scholar
  7. 7.
    Ward PS, Patel J, Wise DR, et al. The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate. Cancer Cell. 2010;17(3):225–34.CrossRefGoogle Scholar
  8. 8.
    Gross S, Cairns RA, Minden MD, et al. Cancer-associated metabolite 2-hydroxyglutarate accumulates in acute myelogenous leukemia with isocitrate dehydrogenase 1 and 2 mutations. J Exp Med. 2010;207(2):339–44.CrossRefGoogle Scholar
  9. 9.
    Yan H, Parsons DW, Jin G, et al. IDH1 and IDH2 mutations in gliomas. N Engl J Med. 2009;360(8):765–73.CrossRefGoogle Scholar
  10. 10.
    Parsons DW, Jones S, Zhang X, et al. An integrated genomic analysis of human glioblastoma multiforme. Science. 2008;321(5897):1807–12.CrossRefGoogle Scholar
  11. 11.
    Amary MF, Bacsi K, Maggiani F, et al. IDH1 and IDH2 mutations are frequent events in central chondrosarcoma and central and periosteal chondromas but not in other mesenchymal tumours. J Pathol. 2011;224(3):334–43.CrossRefGoogle Scholar
  12. 12.
    Borger DR, Tanabe KK, Fan KC, et al. Frequent mutation of isocitrate dehydrogenase (IDH)1 and IDH2 in cholangiocarcinoma identified through broad-based tumor genotyping. Oncologist. 2012;17(1):72–9.CrossRefGoogle Scholar
  13. 13.
    Mardis ER, Ding L, Dooling DJ, et al. Recurring mutations found by sequencing an acute myeloid leukemia genome. N Engl J Med. 2009;361(11):1058–66.CrossRefGoogle Scholar
  14. 14.
    Marcucci G, Maharry K, Wu YZ, et al. IDH1 and IDH2 gene mutations identify novel molecular subsets within de novo cytogenetically normal acute myeloid leukemia: a Cancer and Leukemia Group B study. J Clin Oncol. 2010;28(14):2348–55.CrossRefGoogle Scholar
  15. 15.
    Cerami E, Gao J, Dogrusoz U, et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2012;2(5):401–4.CrossRefGoogle Scholar
  16. 16.
    Lu J, Xu L, Zou Y, et al. IDH1 p.R132 mutations may not be actively involved in the carcinogenesis of hepatocellular carcinoma. Med Sci Monit. 2014;20:247–54.CrossRefGoogle Scholar
  17. 17.
    Li WL, Xiao MS, Zhang DF, et al. Mutation and expression analysis of the IDH1, IDH2, DNMT3A, and MYD88 genes in colorectal cancer. Gene. 2014;546(2):263–70.CrossRefGoogle Scholar
  18. 18.
    Koivunen P, Lee S, Duncan CG, et al. Transformation by the (R)-enantiomer of 2-hydroxyglutarate linked to EGLN activation. Nature. 2012;483(7390):484–8.CrossRefGoogle Scholar
  19. 19.
    Chowdhury R, Yeoh KK, Tian YM, et al. The oncometabolite 2-hydroxyglutarate inhibits histone lysine demethylases. EMBO Rep. 2011;12(5):463–9.CrossRefGoogle Scholar
  20. 20.
    Xu W, Yang H, Liu Y, et al. Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of alpha-ketoglutarate-dependent dioxygenases. Cancer Cell. 2011;19(1):17–30.CrossRefGoogle Scholar
  21. 21.
    Iwagami S, Baba Y, Watanabe M, et al. Pyrosequencing assay to measure LINE-1 methylation level in esophageal squamous cell carcinoma. Ann Surg Oncol. 2012;19(8):2726–32.CrossRefGoogle Scholar
  22. 22.
    Iwagami S, Baba Y, Watanabe M, et al. LINE-1 hypomethylation is associated with a poor prognosis among patients with curatively resected esophageal squamous cell carcinoma. Ann Surg. 2013;257(3):449–55.CrossRefGoogle Scholar
  23. 23.
    Niibori-Nambu A, Midorikawa U, Mizuguchi S, et al. Glioma initiating cells form a differentiation niche via the induction of extracellular matrices and integrin alphaV. PLoS ONE. 2013;8(5):e59558.CrossRefGoogle Scholar
  24. 24.
    Matsunaga H, Futakuchi-Tsuchida A, Takahashi M, Ishikawa T, Tsuji M, Ando O. IDH1 and IDH2 have critical roles in 2-hydroxyglutarate production in D-2-hydroxyglutarate dehydrogenase depleted cells. Biochem Biophys Res Commun. 2012;423(3):553–6.CrossRefGoogle Scholar
  25. 25.
    Struys EA. 2-Hydroxyglutarate is not a metabolite; D-2-hydroxyglutarate and L-2-hydroxyglutarate are! Proc. Natl. Acad. Sci. USA. 2013;110(51):E4939.CrossRefGoogle Scholar
  26. 26.
    Kranendijk M, Struys EA, Salomons GS, Van der Knaap MS, Jakobs C. Progress in understanding 2-hydroxyglutaric acidurias. J Inherit Metab Dis. 2012;35(4):571–87.CrossRefGoogle Scholar
  27. 27.
    Cairns RA, Mak TW. Oncogenic isocitrate dehydrogenase mutations: mechanisms, models, and clinical opportunities. Cancer Discov. 2013;3(7):730–41.CrossRefGoogle Scholar
  28. 28.
    Fujii T, Khawaja MR, DiNardo CD, Atkins JT, Janku F. Targeting isocitrate dehydrogenase (IDH) in cancer. Discov Med. 2016;21(117):373–80.PubMedGoogle Scholar
  29. 29.
    Zhao G, Winkler ME. A novel alpha-ketoglutarate reductase activity of the serA-encoded 3-phosphoglycerate dehydrogenase of Escherichia coli K-12 and its possible implications for human 2-hydroxyglutaric aciduria. J Bacteriol. 1996;178(1):232–9.CrossRefGoogle Scholar
  30. 30.
    Achouri Y, Noel G, Vertommen D, Rider MH, Veiga-Da-Cunha M, Van Schaftingen E. Identification of a dehydrogenase acting on D-2-hydroxyglutarate. Biochem J. 2004;381(Pt 1):35–42.CrossRefGoogle Scholar
  31. 31.
    Linster CL, Van Schaftingen E, Hanson AD. Metabolite damage and its repair or pre-emption. Nat Chem Biol. 2013;9(2):72–80.CrossRefGoogle Scholar
  32. 32.
    Intlekofer AM, Dematteo RG, Venneti S, et al. Hypoxia Induces Production of L-2-Hydroxyglutarate. Cell Metab. 2015;22(2):304–11.CrossRefGoogle Scholar
  33. 33.
    Rzem R, Van Schaftingen E, Veiga-da-Cunha M. The gene mutated in l-2-hydroxyglutaric aciduria encodes l-2-hydroxyglutarate dehydrogenase. Biochimie. 2006;88(1):113–6.CrossRefGoogle Scholar
  34. 34.
    Shim EH, Livi CB, Rakheja D, et al. L-2-Hydroxyglutarate: an epigenetic modifier and putative oncometabolite in renal cancer. Cancer Discov. 2014;4(11):1290–8.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Keisuke Miyake
    • 1
  • Yoshifumi Baba
    • 1
  • Takatsugu Ishimoto
    • 1
  • Yukiharu Hiyoshi
    • 1
  • Masaaki Iwatsuki
    • 1
  • Yuji Miyamoto
    • 1
  • Naoya Yoshida
    • 1
  • Masayuki Watanabe
    • 2
  • Yoko Ogata
    • 1
  • Megumi Nagayama
    • 3
  • Atit Silsirivanit
    • 3
    • 4
  • Daiki Kobayashi
    • 3
  • Norie Araki
    • 3
  • Hideo Baba
    • 1
    Email author
  1. 1.Department of Gastroenterological Surgery, Graduate School of Medical ScienceKumamoto UniversityKumamotoJapan
  2. 2.Department of Gastroenterological Surgery, Cancer Institute HospitalJapanese Foundation for Cancer ResearchTokyoJapan
  3. 3.Department of Tumor Genetics and Biology, Graduate School of Medical SciencesKumamoto UniversityKumamotoJapan
  4. 4.Department of Biochemistry, Faculty of Medicine, Cholangiocarcinoma Research CenterKhon Kaen UniversityKhon KaenThailand

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