Genes & Nutrition

, Volume 2, Issue 2, pp 225–232 | Cite as

Evidence that a polymorphism within the 3′UTR of glutathione peroxidase 4 is functional and is associated with susceptibility to colorectal cancer

  • G. Bermano
  • V. Pagmantidis
  • N. Holloway
  • S. Kadri
  • N. A. G. Mowat
  • R. S. Shiel
  • J. R. Arthur
  • J. C. Mathers
  • A. K. Daly
  • J. Broom
  • J. E. HeskethEmail author
Research Paper


Low selenium (Se) status has been associated with increased risk of colorectal cancer (CRC). Se is present as the amino acid selenocysteine in selenoproteins, such as the glutathione peroxidases. Se incorporation requires specific RNA structures in the 3′ untranslated region (3′UTR) of the selenoprotein mRNAs. A single nucleotide polymorphism (SNP) occurs at nucleotide 718 (within the 3′UTR) in the glutathione peroxidase 4 gene. In the present study, Caco-2 cells were transfected with constructs in which type 1 iodothyronine deiodinase coding region was linked to the GPx4 3′UTR with either C or T variant at position 718. Higher reporter activity was observed in cells expressing the C variant compared to those expressing the T variant, under either Se-adequate or Se-deficient conditions. In addition, a disease association study was carried out in cohorts of patients with either adenomatous polyps, colorectal adenocarcinomas and in healthy controls. A higher proportion of individuals with CC genotype at the GPx4 T/C 718 SNP was present in the cancer group, but not in the polyp group, compared with the control group (P < 0.05). The present data demonstrate the functionality of the GPx4 T/C 718 SNP and suggest that T genotype is associated with lower risk of CRC.


Colorectal cancer GPx4 Reporter gene Selenium SNP 3′Untranslated region 



We thank the World Cancer Research Fund for financial support (grants 2000/10, 2002/41), clinical staff for assistance with sampling and the patients for consenting to take part in this study. We greatly acknowledge Brian Burtle, Rebecca Lamb, Leanne Boulding and Rachel Ledward for help with genotyping and Fergus Nicol for help with IDI activity assay. JRA’s lab is funded by The Scottish Executive Environment and Rural Affairs Department (SEERAD). Conflict of interest-None declared.


  1. 1.
    Akbaraly NT, Arnaud J, Hininger-Favier I, Gourlet V, Roussel AM, Berr C (2005) Selenium and mortality in the elderly: results from the EVA study. Clin Chem 51:2117–2123PubMedCrossRefGoogle Scholar
  2. 2.
    Arthur JR, Nicol F, Beckett GJ (1990) Hepatic type 1 iodothyronine 5′-deiodinase. The role of selenium. Biochem J 272:537–540PubMedGoogle Scholar
  3. 3.
    Bermano G, Nicol F, Dyer JA, Sunde RA, Beckett GJ, Arthur JR, Hesketh JE (1995) Tissue-specific regulation of selenoenzyme gene expression during selenium deficiency in rats. Biochem J 311:425–430PubMedGoogle Scholar
  4. 4.
    Bermano G, Arthur JR, Hesketh JE (1996) Role of the 3′ untranslated region in the regulation of cytosolic glutathione peroxidase and phospholipid-hydroperoxide glutathione peroxidase gene expression by selenium supply. Biochem J 320:891–895PubMedGoogle Scholar
  5. 6.
    Berry MJ (2005) Insights into the hierarchy of selenium incorporation. Nat Genet 37:1162–1163PubMedCrossRefGoogle Scholar
  6. 5.
    Berry MJ, Banu L, Chen YY, Mandel SJ, Kieffer JD, Harney JW, Larsen PR (1991) Recognition of UGA as a selenocysteine codon in type I deiodinase requires sequences in the 3′ untranslated region. Nature 353:273–276PubMedCrossRefGoogle Scholar
  7. 8.
    Brigelius-Flohe R (1999) Tissue-specific functions of individual glutathione peroxidases. Free Radic Biol Med 27:951–965PubMedCrossRefGoogle Scholar
  8. 7.
    Brigelius-Flohe R, Friedrichs B, Maurer S, Schultz M, streicher R (1997) Interleukin-1-induced nuclear factor kappa B activation is inhibited by overexpression of phospholipid hydroperoxide glutathione peroxidase in a human endothelial cell line. Biochem J 328:199–203PubMedGoogle Scholar
  9. 9.
    Clark LC, Hixson LJ, Combs GF, Reid ME, Turnbull BW, Sampliner RE (1993) Plasma selenium concentration predicts the prevalence of colorectal adenomatous polyps. Cancer Epidemiol Biomark Prev 2:41–46Google Scholar
  10. 10.
    Clark LC, Combs GF Jr, Turnbull BW, Slate EH, Chalker DK, Chow J, Davis LS, Glover RA, Graham GF, Gross EG, Krongrad A, Lesher JL Jr, Park HK, Sanders BB Jr, Smith CL, Taylor JR (1996) Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. A randomized controlled trial. Nutritional Prevention of Cancer Study Group. J Am Med Assoc 276:1957–1963CrossRefGoogle Scholar
  11. 12.
    Combs GF (2005) Current evidence and research needs to support a health claim for selenium and cancer prevention. J Nutr 135:343–347PubMedGoogle Scholar
  12. 11.
    Combs GF Jr, Gray WP (1998) Chemopreventive agents: selenium. Pharmacol Ther 79:179–192PubMedCrossRefGoogle Scholar
  13. 13.
    Cummings OW (2000) Pathology of the adenoma–carcinoma sequence: from aberrant crypt focus to invasive carcinoma. Semin Gastrointest Dis 11:229–237PubMedGoogle Scholar
  14. 14.
    Diplock AT (1993) Indexes of selenium status in human populations. Am J Clin Nutr 57:256S–258SPubMedGoogle Scholar
  15. 15.
    Dworkin BM, Rosenthal WS, Mittelman A, Weiss L, Applebee-Brady L, Arlin Z (1988) Selenium status and the polyp-cancer sequence: a colonoscopy controlled study. Am J Gastroenterol 83:748–751PubMedGoogle Scholar
  16. 16.
    Esworthy RS, Aranda R, Martin MG, Doroshow JH, Binder SW, Chu FF (2001) Mice with combined disruption of Gpx1 and Gpx2 genes have colitis. Am J Physiol Gastrointest Liver Physiol 281:G848–55PubMedGoogle Scholar
  17. 17.
    Esworthy RS, Yang L, Frankel PH, Chu FF (2005) Epithelium-specific glutathione peroxidase, Gpx2, is involved in the prevention of intestinal inflammation in selenium-deficient mice. J Nutr 135:740–745PubMedGoogle Scholar
  18. 18.
    Forsberg L, de Faire U, Morgenstern R (1998) To identify genetic polymorphisms in the “Expressed Sequence Tag” (EST) database. Technical tips on-line http://www.elsevier.com_/locate_/tto, T01440
  19. 19.
    Forsberg L, de Faire U, Marklund SL, Andersson PM, Stegmayr B, Morgenstern R (2000) Phenotype determination of a common Pro-Leu polymorphism in human glutathione peroxidase 1. Blood Cells Mol Dis 26:423–426PubMedCrossRefGoogle Scholar
  20. 20.
    Haklar G, Sayin-Ozveri E, Yuksel M, Aktan AO, Yalcin AS (2001) Different kinds of reactive oxygen and nitrogen species were detected in colon and breast tumors. Cancer Lett 165:219–224PubMedCrossRefGoogle Scholar
  21. 21.
    Heirman I, Ginneberge D, Brigelius-Flohe R, Hendrickx N, Agostinis P, Brouckaert P, Rottiers P. And Grooten J (2006) Blocking tumor cell eicosanoid synthesis by GPx4 impedes tumor growth and malignancy. Free Radic Biol Med 40:285–94PubMedCrossRefGoogle Scholar
  22. 23.
    Hu YJ, Diamond AM (2003) Role of glutathione peroxidase 1 in breast cancer: loss of heterozygosity and allelic differences in the response to selenium. Cancer Res 63:3347–3351PubMedGoogle Scholar
  23. 22.
    Hu YJ, Korotkov KV, Mehta R, Hatfield DL, Rotimi CN, Luke A, Prewitt TE, Cooper RS, Stock W, Vokes EE, Dolan ME, Gladyshev VN, Diamond AM (2001) Distribution and functional consequences of nucleotide polymorphisms in the 3′-untranslated region of the human Sep15 gene. Cancer Res 61:2307–2310PubMedGoogle Scholar
  24. 24.
    Ichimura Y, Habuchi T, Tsuchiya N, Wang L, Oyama C, Sato K, Nishiyama H, Ogawa O, Kato T (2004) Increased risk of bladder cancer associated with a glutathione peroxidase 1 codon 198 variant. J Urol 172:728–732PubMedCrossRefGoogle Scholar
  25. 25.
    Imai H, Nakagawa Y (2003) Biological significance of phospholipid hydroperoxide glutathione peroxidase (PHGPx, GPx4) in mammalian cells. Free Radic Biol Med 34:145–169PubMedCrossRefGoogle Scholar
  26. 26.
    Itzkowitz SH, Yio X (2004) Inflammation and cancer IV. Colorectal cancer in inflammatory bowel disease: the role of inflammation. Am J Physiol Gastrointest Liver Physiol 287:G7–G17PubMedCrossRefGoogle Scholar
  27. 27.
    Jacobs ET, Jiang R, Alberts DS, Greenberg ER, Gunter EW, Karagas MR, Lanza E, Ratnasinghe L, Reid ME, Schatzkin A, Smith-Warner SA, Wallace K, Martinez ME (2004) Selenium and colorectal adenoma: results of a pooled analysis. J Natl Cancer Instit 96:1669–1675CrossRefGoogle Scholar
  28. 28.
    Jacobson-Brown P, Neuman MG (2004) Colorectal polyposis and immune-based therapies. Canad J Gastroenterol 18:239–249Google Scholar
  29. 29.
    Kollmus H, Flohe L, McCarthy JE (1996) Analysis of eukaryotic mRNA structures directing cotranslational incorporation of selenocysteine. Nucleic Acids Res 24:1195–1201PubMedCrossRefGoogle Scholar
  30. 30.
    Kryukov GV, Kryukov VM, Gladyshev VN (1999) New mammalian selenocysteine-containing proteins identified with an algorithm that searches for selenocysteine insertion sequence elements. J Biol Chem 274:33888–33897PubMedCrossRefGoogle Scholar
  31. 31.
    Nomura K, Imai H, Koumura T, Arai M, Nakagawa Y (1999) Mitochondrial phospholipid hydroperoxide glutathione peroxidase suppresses apoptosis mediated by a mitochondrial death pathway. J Biol Chem 274:22294–29302CrossRefGoogle Scholar
  32. 32.
    Pagmantidis V, Bermano G, Villette S, Broom J, Arthur J, Hesketh JE (2005) Effects of Se-depletion on glutathione peroxidase and SelW gene expression in the colon. FEBS Lett 579:792–796PubMedCrossRefGoogle Scholar
  33. 33.
    Qatatsheh A, Seal CJ, Jowett SL, Welfare MR, Hesketh JE (2005) Patients with ulcerative colitis show an altered frequency distribution of a single nucleotide polymorphism in the gene encoding the phospholipid hydroperoxide glutathione peroxidase. Proceedings of the Nutrition Society 64, OCA, 20A (Abstract)Google Scholar
  34. 34.
    Rayman MP (2005) Selenium in cancer prevention: a review of the evidence and mechanism of action. Proc Nutr Soc 64:527–542PubMedCrossRefGoogle Scholar
  35. 35.
    Russo MW, Murray SC, Wurzelmann JI, Woosley JT, Sandler RS (1997) Plasma selenium levels and the risk of colorectal adenomas. Nutr Cancer 28:125–129PubMedCrossRefGoogle Scholar
  36. 36.
    Rutter M, Saunders B, Wilkinson K, Rumbles S, Schofield G, Kamm M, Williams C, Price A, Talbot I, Forbes A (2004) Severity of inflammation is a risk factor for colorectal neoplasia in ulcerative colitis. Gastroenterology 126:451–459PubMedCrossRefGoogle Scholar
  37. 37.
    Schnurr K, Belkner J, Ursini F, Schewe T, Kuhn H (1996) The selenoenzyme phospholipid hydroperoxide glutathione peroxidase controls the activity of the 15-lipoxygenase with complex substrates and preserves the specificity of the oxygenation products. J Biol Chem 271:4653–4658PubMedCrossRefGoogle Scholar
  38. 38.
    Seril DN, Liao J, Yang GY, Yang CS (2003) Oxidative stress and ulcerative colitis-associated carcinogenesis: studies in humans and animal models. Carcinogenesis 24:353–362PubMedCrossRefGoogle Scholar
  39. 39.
    Shamberger RJ, Tytko SA, Willis CE (1976) Antioxidants and cancer. Part VI. Selenium and age-adjusted human cancer mortality. Arch Environ Health 31:231–235PubMedGoogle Scholar
  40. 40.
    Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchoninic acid. Ann Biochem 150:76–85CrossRefGoogle Scholar
  41. 41.
    Stadtman TC (1996) Selenocysteine. Ann Rev Biochem 65:83–100PubMedCrossRefGoogle Scholar
  42. 42.
    Villette S, Kyle JA, Brown KM, Pickard K, Milne JS, Nicol F, Arthur JR, Hesketh JE (2002) A novel single nucleotide polymorphism in the 3′ untranslated region of human glutathione peroxidase 4 influences lipoxygenase metabolism. Blood Cells Mol Dis 29:174–178PubMedCrossRefGoogle Scholar
  43. 43.
    Whanger PD (2004) Selenium and its relationship to cancer: an update dagger. Br J Nutr 91:11–28PubMedCrossRefGoogle Scholar
  44. 44.
    Wingler K, Böcher M, Flohe L, Kollmus H, Brigelius-Flohe R (1999) mRNA stability and selenocysteine insertion sequence efficiency rank gastrointestinal glutathione peroxidase high in the hierarchy of selenoproteins. Eur J Biochem 259:149–157PubMedCrossRefGoogle Scholar
  45. 45.
    Yagi K, Komura S, Kojima H, Sun Q, Nagata N, Ohishi N, Nishikimi M (1996) Expression of human phospholipid hydroperoxide glutathione peroxidase gene for protection of host cells from lipid hydroperoxide-mediated injury. Biochem Biophys Res Commun 219:486–491PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • G. Bermano
    • 1
  • V. Pagmantidis
    • 2
  • N. Holloway
    • 1
  • S. Kadri
    • 3
  • N. A. G. Mowat
    • 3
  • R. S. Shiel
    • 4
  • J. R. Arthur
    • 5
  • J. C. Mathers
    • 6
  • A. K. Daly
    • 8
  • J. Broom
    • 1
  • J. E. Hesketh
    • 7
    Email author
  1. 1.School of Life SciencesThe Robert Gordon UniversityAberdeenUK
  2. 2.Institute for Cell and Molecular BiosciencesNewcastle UniversityNewcastle upon TyneUK
  3. 3.Aberdeen Royal InfirmaryAberdeenUK
  4. 4.School of Agriculture and Rural DevelopmentNewcastle UniversityNewcastle upon TyneUK
  5. 5.Rowett Research InstituteAberdeenUK
  6. 6.School of Clinical Medical Sciences and Human Nutrition Research CentreNewcastle UniversityNewcastle upon TyneUK 
  7. 7.Institute for Cell and Molecular Biosciences and Human Nutrition Research CentreNewcastle UniversityNewcastle upon TyneUK 
  8. 8.School of Clinical and Laboratory SciencesNewcastle UniversityNewcastle upon TyneUK 

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