Staging of colorectal cancer using serum metabolomics with 1HNMR Spectroscopy


1 Biochemistry Department, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran

2 Amir Alam Hospital, Amir Alam, Tehran, Iran

3 Payam Noor University, Tehran, Iran


Objective(s): Determination of stages of colon cancer is done by biopsy usually after surgery. Metabolomics is the study of all the metabolites using LC-MS and 1HNMR spectroscopy with chemometric techniques. The stages of colon cancer were detected from patients' sera using 1HNMR.
Materials and Methods: Five ml blood was collected from 16 confirmed patients referred for colonoscopy.  One group of eight patients were diagnosed with stage 0 to I colon cancer and the second group of 8 patients with II-IV stage colon cancer.  Sera were sent for 1HNMR. The differentiating metabolites were identified using HMDB  and the metabolic cycles from Metaboanalyst.
Results: Six metabolites of which pyridoxine levels lowered, and glycine, cholesterol, taurocholic acid, cholesteryl ester and deoxyinosine increased.
Conclusion: The different stages of cancer were identified by the main metabolic cycles such as primary bile acid biosynthesis, purine and vitamin B metabolic pathways and the glutathione cycle.


1. Seer Stat Fact Sheets. Colon and Rectum Cancer National cancer institute. Available at:
2. Safaee A, Fatemi SR, Ashtari S, Vahedi M, Moghimi-Dehkordi B. Four years incidence rate of colorectal cancer in Iran. a survey of national cancer registry data implications for screening. Asian Pac J Cancer Prev 2012; 13:26952-2698.
3. Ansari R, Mahdavinia M, Sadjadi A, Nouraie M, Kamangar F, Bishehsari F, et al. Incidence and age distribution of colorectal cancer in Iran results of a population-based cancer registry. Cancer Lett 2006; 240:143-147.
4. Colon Cancer Treatment (PDQ®)-National cancer institute. Available at:    cancertopics/pdq/treatment/colon/Patient/p.2.
5. Nicholson JK, Lindon JC, Holmes E. Metabonomics: understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica1999; 12:1181–1189.
6. Fiehn O. Metabolomics-the link between genotypes and phenotypes. Plant Mol Biol 2002; 12:155–171.
7. Jeremy K, Connelly NG, Lindon,JC, Holmes E. Metabonomics:a platform for studying drug toxicity and gene function. Nat Rev Drug Discov 2002; 12:153–162.
8. Claudino WM, Quattrone A, Biganzoli L, QuattronA, Pestrin M, Bertini I, et al. Metabolomics: available results, current research projects in breast cancer, and future applications. J Clin Oncol 2007; 12:2840–2846.
9. Xue R, Lin Z, Deng C, Dong L, Liu T, Wang J, et al. Serum metabolomics investigation on hepatocellular carcinoma patients by chemical derivatization followed by gas chromatography/mass spectrometry. Rapid Commun Mass Spectrom 2008; 12:3061–3068.
10.  Wei L, Liao P, Wu H, Li X, Pei F, Li W, et al. Toxicological effects of cinnabar in rats by NMR-based metabolic profiling of urine and serum. Toxicol Appl Pharm 2008; 12:417–429.
11. Van Asten JJ, Cuijpers V, Hulsbergen-van De Kaa C, Soede-Huijbregts C, Witjes JA, Verhofstad A, et al. High-resolution magic angle spinning NMR spectroscopy for metabolic assessment of cancer presence and Gleason score in human prostate needle biopsies. MAGMA 2008; 12:435–442.
12. Astrakas LG, Zurakowski D, Tzika AA, Zarifi MK, Anthony DC, De Girolami U, et al. Noninvasive magnetic resonance spectroscopic imaging biomarkers to predict the clinical grade of pediatric brain tumors. Clin Cancer Res 2004; 12:8220–8228.
13. Jagannathan NP and Sharma U. Breast tissue metabolism by magnetic resonance spectroscopy metabolites 2017, 7, 25; doi:10.3390/metabo7020025.
14. Wishart DS, Jewison T, Guo AC, Wilson M, Knox C, Liu Y, et al .HMDB 3.0 — The Human resonance spectroscopy. Metabolomics 2009; 12:292–301.
15. Farshidfar F, Weljie AM, Kopciuk KA, Hilsden R, McGregor SE, Buie WD, et al.  A validated metabolomic signature for colorectal cancer: exploration of the clinical value of metabolomics. Br J Cancer 2016; 115:848-857. 
16. Xia J, Mandal R, Sinelnkoviv, Broadhurst D , Wishart D. MetaboAnalyst 2.0--a comprehensive server for metabolomic data analysis. Nucleic Acids Res 2012l; 40.
17. Zamani Z, Arjmand M,  Vahabi F. IshaqHosseini SM, Fazeli SM, Sadeghi S, et al. A Metabolic study on colon cancer using 1H nuclear magnetic resonance spectroscopy. Biochem Res Int 2014; 2014:348712.
18. Seidel A, Seidel P, Manuwald O.  Modified nucleosides as biomarkers for early cancer diagnose in exposed populations.Environ Toxicol 2015; 30:956-967.
19. Hsu WY, Chen CJ, Huang YC, Tsai FJ,  Jeng  LB,  Lai CC. Urinary nucleosides as biomarkers of breast, colon, lung, and gastric cancer in Taiwanese.  PLoS One 2013; 8:e81701.
20. Dustin G. Brown, Sangeeta Rao, Tiffany L. Weir, Joanne O’Malia, Marlon Bazan, Regina J. Brown, and Elizabeth P. Ryan Metabolomics and metabolic pathway networks from human colorectal cancers, adjacent mucosa, and stoolCancer Metab. 2016; 4: 11. Published online 2016 Jun 6. doi:  10.1186/s40170-016-0151-y
21. Komatsu S, Yanaka N, Matsubara K, Kato N. Antitumor effect of vitamin B6 and its mechanisms. Biochim Biophys Acta 2003; 1647:127-130. 
22. Zhang P, Suidasari S, Hasegawa T,  Yanaka N, Kato N. Vitamin. B₆ activates p53 and elevates p21 gene expression in cancer cells and the mouse colon. Oncol Rep 2014; 31:2371-2376.
23. Kayashima T, Tanaka K, Okazaki Y,  Matsubara K, Yanaka N, Kato  N. Consumption of vitamin B6 reduces colonic damage and protein expression of HSP70 and HO-1, the anti-tumor targets, in rats exposed to 1,2-dimethylhydrazine.  Oncol Lett 2011; 2:1243-1246.
24.  Ajouz  H, Mukherji D ,Shamseddine A. Secondary bile acids: an under-recognized cause of colon cancer.  World J Surg Oncol 2014; 12:164.
25. Payne CM, Bernstein C, Dvorak K. Hydrophobic bile acids, genomic instability, Darwinian selection, and colon carcinogenesis. Clin Exp Gastroenterol 2008; 1:19-47.
26. Okazaki Y, Utama Z, Suidasari S, Zhang P, Tanaka N, Tomotake H, et al. Consumption of vitamin B(6) reduces fecal ratio of lithocholic acid to deoxycholic acid, a risk factor for colon cancer, in rats fed a high-fat diet. J Nutr Sci Vitaminol 2012; 5:366-370.
27. Bayerdörffer E, Mannes GA, Richter WO, Ochsenkühn T, Wiebecke B,  Köpcke W, et al. Increased serum deoxycholic acid levels in men with colorectal adenomas. Gastroentero-logy 1993; 104:145-151.
28.  Jain M,  Nilsson R,  Sharma S,  Madhusudhan N,  Kitami T, Souza AL, et al. Metabolite profiling identifies a key role for glycine in rapid cancer cell proliferation. Science 2012; 336 :1040.
29.  Locasale JW. Serine, glycine and one-carbon units: cancer metabolism in a full circle. Nat Rev Cancer Aug 2013; 13:572–583.
30. Zhang WC, Shih-Chang N, Yang H, Rai A, Umashankar S, Ma S, et al. Glycine decarboxylase activity drives non-small cell lung cancer tumor-initiating cells and tumorigenesis. Cell 2012; 148:259-272.
31. Paterson AC, Macrae FA, Pizzey C, Baldwin GS, Shulkes A. Circulating gastrin concentrations in patients at increased risk of developing colorectal carcinoma. J Gastroenterol Hepatol 2014; 29:480-486.
32. Tour S, Sugiura Y, Kubo A, Ohmura M, Karakawa S, Mizukoshi T. Microscopic imaging mass spectrometry assisted by on-tissue chemical derivatization for visualizing multiple amino acids in human colon cancer xenografts. Proteomics 2014; 14:810-819.
33. Andreyev HJ, Norman AR, Cunningham D, Oates J, Dix BR, Iacopetta BJ, et al. Kirsten ras mutations in patients with colorectal cancer: the 'RASCAL II' study.  J Cancer 2001; 85:692-696.
34.  Grubben MJ, Nagengast FM, Katan MB, Peters WH. The glutathione biotransformation system and colorectal cancer risk in humans. Scand J Gastroenterol Suppl 2001; 68-76.
35. Haug U, Poole EM, Xiao L, Curtin K, Duggan D, Hsu L,
et al. Glutathione peroxidase tagSNPs: Associations with rectal cancer but not with colon cancer. Genes Chromosomes Cancer 2012; 51:598-605.
36. Jankova L, Robertson G, Chan C, Tan KL, Kohonen-Corish M, Fung C L-S, et al. Glutathione S-transferase Pi expression predicts response to adjuvant chemotherapy for stage C colon cancer: a matched historical control study. BMC Cancer 2012; 12:196.