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Evaluation of UDP-glucuronosyltransferase 2B17 (UGT2B17) and dihydrofolate reductase (DHFR) genes deletion and the expression level of NGX6 mRNA in breast cancer

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Abstract

The present study was aimed to investigate the possible association between 19-base pair (bp) deletion polymorphism of the DHFR gene (rs70991108), null genotype of UDP-glucuronosyltransferase 2B17 (UGT2B17) as well as the expression level of nasopharyngeal carcinoma-associated gene 6 (NGX6) with the risk of breast cancer. This case–control study was done on 236 patients with breast cancer and 203 cancer free women. Detection of 19-bp del of DHFR was done using bi-directional PCR allele-specific amplification and UGT2B17 genotyping was performed using multiplex PCR assay. NGX6 mRNA expression level was determined by quantitative reverse transcriptase PCR in 62 breast cancerous and 62 adjacent non-cancerous tissues. Our finding showed an association between null genotype of UGT2B17 and risk of breast cancer and the null genotype increased susceptibility to breast cancer (OR: 2.99; 95 % CI: 1.94–4.60; p < 0.0001). However, no statistically significant difference was found between breast cancer patients and cancer free normal women regarding 19-bp ins/del of DHFR (χ2 = 0.91, p = 0.63). Real-time PCR data showed that the relative expression level of NGX6 mRNA was significantly lower in cancerous than that in non-cancerous breast tissue specimens (0.936 ± 0.042 and 1.042 ± 0.039, respectively). However, NGX6 mRNA expression was not correlated with tumors grade (p > 0.05). In conclusion, the null genotype of UGT2B17 revealed to be a risk factor for breast cancer in a sample of Iranian population. Furthermore, down-regulation of NGX6 mRNA expression in breast carcinoma confirms the growing proof regarding the tumor suppressor role of NGX6.

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References

  1. Key TJ, Verkasalo PK, Banks E (2001) Epidemiology of breast cancer. Lancet Oncol 2:133–140

    Article  PubMed  CAS  Google Scholar 

  2. Cebrian A, Pharoah PD, Ahmed S, Ropero S, Fraga MF, Smith PL, Conroy D, Luben R, Perkins B, Easton DF, Dunning AM, Esteller M, Ponder BA (2006) Genetic variants in epigenetic genes and breast cancer risk. Carcinogenesis 27:1661–1669

    Article  PubMed  CAS  Google Scholar 

  3. Bombonati A, Sgroi DC (2011) The molecular pathology of breast cancer progression. J Pathol 223:307–317

    Article  PubMed  CAS  Google Scholar 

  4. Guillemette C (2003) Pharmacogenomics of human UDP-glucuronosyltransferase enzymes. Pharmacogenomics J 3:136–158

    Article  PubMed  CAS  Google Scholar 

  5. Nagar S, Remmel RP (2006) Uridine diphosphoglucuronosyltransferase pharmacogenetics and cancer. Oncogene 25:1659–1672

    Article  PubMed  CAS  Google Scholar 

  6. Haakensen VD, Biong M, Lingjaerde OC, Holmen MM, Frantzen JO, Chen Y, Navjord D, Romundstad L, Luders T, Bukholm IK, Solvang HK, Kristensen VN, Ursin G, Borresen-Dale AL, Helland A (2010) Expression levels of uridine 5′-diphospho-glucuronosyltransferase genes in breast tissue from healthy women are associated with mammographic density. Breast Cancer Res 12:R65

    Article  PubMed  Google Scholar 

  7. He X, Hesse LM, Hazarika S, Masse G, Harmatz JS, Greenblatt DJ, Court MH (2009) Evidence for oxazepam as an in vivo probe of UGT2B15: oxazepam clearance is reduced by UGT2B15 D85Y polymorphism but unaffected by UGT2B17 deletion. Br J Clin Pharmacol 68:721–730

    Article  PubMed  CAS  Google Scholar 

  8. Hirata H, Hinoda Y, Zaman MS, Chen Y, Ueno K, Majid S, Tripsas C, Rubin M, Chen LM, Dahiya R (2010) Function of UDP-glucuronosyltransferase 2B17 (UGT2B17) is involved in endometrial cancer. Carcinogenesis 31:1620–1626

    Article  PubMed  CAS  Google Scholar 

  9. Hum DW, Belanger A, Levesque E, Barbier O, Beaulieu M, Albert C, Vallee M, Guillemette C, Tchernof A, Turgeon D, Dubois S (1999) Characterization of UDP-glucuronosyltransferases active on steroid hormones. J Steroid Biochem Mol Biol 69:413–423

    Article  PubMed  CAS  Google Scholar 

  10. Park J, Chen L, Ratnashinge L, Sellers TA, Tanner JP, Lee JH, Dossett N, Lang N, Kadlubar FF, Ambrosone CB, Zachariah B, Heysek RV, Patterson S, Pow-Sang J (2006) Deletion polymorphism of UDP-glucuronosyltransferase 2B17 and risk of prostate cancer in African American and Caucasian men. Cancer Epidemiol Biomarkers Prev 15:1473–1478

    Article  PubMed  CAS  Google Scholar 

  11. Yong M, Schwartz SM, Atkinson C, Makar KW, Thomas SS, Newton KM, Aiello Bowles EJ, Holt VL, Leisenring WM, Lampe JW (2010) Associations between polymorphisms in glucuronidation and sulfation enzymes and mammographic breast density in premenopausal women in the United States. Cancer Epidemiol Biomarkers Prev 19:537–546

    Article  PubMed  CAS  Google Scholar 

  12. Turgeon D, Carrier JS, Chouinard S, Belanger A (2003) Glucuronidation activity of the UGT2B17 enzyme toward xenobiotics. Drug Metab Dispos 31:670–676

    Article  PubMed  CAS  Google Scholar 

  13. Zheng Z, Fang JL, Lazarus P (2002) Glucuronidation: an important mechanism for detoxification of benzo[a]pyrene metabolites in aerodigestive tract tissues. Drug Metab Dispos 30:397–403

    Article  PubMed  CAS  Google Scholar 

  14. Fang JL, Beland FA, Doerge DR, Wiener D, Guillemette C, Marques MM, Lazarus P (2002) Characterization of benzo(a)pyrene-trans-7,8-dihydrodiol glucuronidation by human tissue microsomes and overexpressed UDP-glucuronosyltransferase enzymes. Cancer Res 62:1978–1986

    PubMed  CAS  Google Scholar 

  15. Gallagher CJ, Muscat JE, Hicks AN, Zheng Y, Dyer AM, Chase GA, Richie J, Lazarus P (2007) The UDP-glucuronosyltransferase 2B17 gene deletion polymorphism: sex-specific association with urinary 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol glucuronidation phenotype and risk for lung cancer. Cancer Epidemiol Biomarkers Prev 16:823–828

    Article  PubMed  CAS  Google Scholar 

  16. Itaaho K, Mackenzie PI, Ikushiro S, Miners JO, Finel M (2008) The configuration of the 17-hydroxy group variably influences the glucuronidation of beta-estradiol and epiestradiol by human UDP-glucuronosyltransferases. Drug Metab Dispos 36:2307–2315

    Article  PubMed  CAS  Google Scholar 

  17. Juul A, Sorensen K, Aksglaede L, Garn I, Rajpert-De Meyts E, Hullstein I, Hemmersbach P, Ottesen AM (2009) A common deletion in the uridine diphosphate glucuronyltransferase (UGT) 2B17 gene is a strong determinant of androgen excretion in healthy pubertal boys. J Clin Endocrinol Metab 94:1005–1011

    Article  PubMed  CAS  Google Scholar 

  18. Swanson C, Mellstrom D, Lorentzon M, Vandenput L, Jakobsson J, Rane A, Karlsson M, Ljunggren O, Smith U, Eriksson AL, Belanger A, Labrie F, Ohlsson C (2007) The uridine diphosphate glucuronosyltransferase 2B15 D85Y and 2B17 deletion polymorphisms predict the glucuronidation pattern of androgens and fat mass in men. J Clin Endocrinol Metab 92:4878–4882

    Article  PubMed  CAS  Google Scholar 

  19. Terakura S, Murata M, Nishida T, Emi N, Akatsuka Y, Riddell SR, Morishima Y, Kodera Y, Naoe T (2005) A UGT2B17-positive donor is a risk factor for higher transplant-related mortality and lower survival after bone marrow transplantation. Br J Haematol 129:221–228

    Article  PubMed  CAS  Google Scholar 

  20. Murata M, Warren EH, Riddell SR (2003) A human minor histocompatibility antigen resulting from differential expression due to a gene deletion. J Exp Med 197:1279–1289

    Article  PubMed  CAS  Google Scholar 

  21. Balliet RM, Chen G, Gallagher CJ, Dellinger RW, Sun D, Lazarus P (2009) Characterization of UGTs active against SAHA and association between SAHA glucuronidation activity phenotype with UGT genotype. Cancer Res 69:2981–2989

    Article  PubMed  CAS  Google Scholar 

  22. Kaeding J, Belanger J, Caron P, Verreault M, Belanger A, Barbier O (2008) Calcitrol (1alpha, 25-dihydroxyvitamin D3) inhibits androgen glucuronidation in prostate cancer cells. Mol Cancer Ther 7:380–390

    Article  PubMed  CAS  Google Scholar 

  23. Sun D, Sharma AK, Dellinger RW, Blevins-Primeau AS, Balliet RM, Chen G, Boyiri T, Amin S, Lazarus P (2007) Glucuronidation of active tamoxifen metabolites by the human UDP glucuronosyltransferases. Drug Metab Dispos 35:2006–2014

    Article  PubMed  CAS  Google Scholar 

  24. Bu R, Gutierrez MI, Al-Rasheed M, Belgaumi A, Bhatia K (2004) Variable drug metabolism genes in Arab population. Pharmacogenomics J 4:260–266

    Article  PubMed  CAS  Google Scholar 

  25. Duthie SJ (2011) Folate and cancer: how DNA damage, repair and methylation impact on colon carcinogenesis. J Inherit Metab Dis 34:101–109

    Article  PubMed  CAS  Google Scholar 

  26. Nordgard SH, Alnaes GI, Hihn B, Lingjaerde OC, Liestol K, Tsalenko A, Sorlie T, Lonning PE, Borresen-Dale AL, Kristensen VN (2008) Pathway based analysis of SNPs with relevance to 5-FU therapy: relation to intratumoral mRNA expression and survival. Int J Cancer 123:577–585

    Article  PubMed  CAS  Google Scholar 

  27. Ongaro A, De Mattei M, Della Porta MG, Rigolin G, Ambrosio C, Di Raimondo F, Pellati A, Masieri FF, Caruso A, Catozzi L, Gemmati D (2009) Gene polymorphisms in folate metabolizing enzymes in adult acute lymphoblastic leukemia: effects on methotrexate-related toxicity and survival. Haematologica 94:1391–1398

    Article  PubMed  CAS  Google Scholar 

  28. Johnson WG, Stenroos ES, Spychala JR, Chatkupt S, Ming SX, Buyske S (2004) New 19 bp deletion polymorphism in intron-1 of dihydrofolate reductase (DHFR): a risk factor for spina bifida acting in mothers during pregnancy? Am J Med Genet A 124A:339–345

    Article  PubMed  Google Scholar 

  29. Xu X, Gammon MD, Wetmur JG, Rao M, Gaudet MM, Teitelbaum SL, Britton JA, Neugut AI, Santella RM, Chen J (2007) A functional 19-base pair deletion polymorphism of dihydrofolate reductase (DHFR) and risk of breast cancer in multivitamin users. Am J Clin Nutr 85:1098–1102

    PubMed  CAS  Google Scholar 

  30. Gemmati D, De Mattei M, Catozzi L, Della Porta M, Serino ML, Ambrosio C, Cuneo A, Friso S, Krampera M, Orioli E, Zeri G, Ongaro A (2009) DHFR 19-bp insertion/deletion polymorphism and MTHFR C677T in adult acute lymphoblastic leukaemia: is the risk reduction due to intracellular folate unbalancing? Am J Hematol 84:526–529

    Article  PubMed  CAS  Google Scholar 

  31. Wang L, Xiang B, Yi M, Zhang WL, Yang JB, Peng SP, Li XL, Li GY (2010) Identification of a new seven-span transmembrane protein: NGX6a is downregulated in nasopharyngeal carcinoma and is associated with tumor metastasis. J Histochem Cytochem 58:41–51

    Article  PubMed  CAS  Google Scholar 

  32. Guo Q, Shen S, Liao M, Lian P, Wang X (2010) NGX6 inhibits cell invasion and adhesion through suppression of Wnt/beta-catenin signal pathway in colon cancer. Acta Biochim Biophys Sin (Shanghai) 42:450–456

    Article  CAS  Google Scholar 

  33. Liu M, Peng Y, Wang X, Guo Q, Shen S, Li G (2010) NGX6 gene mediated by promoter methylation as a potential molecular marker in colorectal cancer. BMC Cancer 10:160

    Article  PubMed  CAS  Google Scholar 

  34. Normanno N, De Luca A, Bianco C, Strizzi L, Mancino M, Maiello MR, Carotenuto A, De Feo G, Caponigro F, Salomon DS (2006) Epidermal growth factor receptor (EGFR) signaling in cancer. Gene 366:2–16

    Article  PubMed  CAS  Google Scholar 

  35. Ma J, Zhou J, Fan S, Wang L, Li X, Yan Q, Zhou M, Liu H, Zhang Q, Zhou H, Gan K, Li Z, Peng C, Xiong W, Tan C, Shen S, Yang J, Li J, Li G (2005) Role of a novel EGF-like domain-containing gene NGX6 in cell adhesion modulation in nasopharyngeal carcinoma cells. Carcinogenesis 26:281–291

    Article  PubMed  CAS  Google Scholar 

  36. Xiao JD, Shen SR (2008) Expression and significance of NGX6 gene in human hepatocellular carcinoma. Zhong Nan Da Xue Xue Bao Yi Xue Ban 33:937–941

    PubMed  CAS  Google Scholar 

  37. Zhang XM, Wang XY, Sheng SR, Wang JR, Li J (2003) Expression of tumor related genes NGX6, NAG-7, BRD7 in gastric and colorectal cancer. World J Gastroenterol 9:1729–1733

    PubMed  CAS  Google Scholar 

  38. Hashemi M, Moazeni-Roodi AK, Fazaeli A, Sandoughi M, Bardestani GR, Kordi-Tamandani DM, Ghavami S (2010) Lack of association between paraoxonase-1 Q192R polymorphism and rheumatoid arthritis in southeast Iran. Genet Mol Res 9:333–339

    Article  PubMed  CAS  Google Scholar 

  39. Pande M, Amos CI, Osterwisch DR, Chen J, Lynch PM, Broaddus R, Frazier ML (2008) Genetic variation in genes for the xenobiotic-metabolizing enzymes CYP1A1, EPHX1, GSTM1, GSTT1, and GSTP1 and susceptibility to colorectal cancer in Lynch syndrome. Cancer Epidemiol Biomarkers Prev 17:2393–2401

    Article  PubMed  CAS  Google Scholar 

  40. Setlur SR, Chen CX, Hossain RR, Ha JS, Van Doren VE, Stenzel B, Steiner E, Oldridge D, Kitabayashi N, Banerjee S, Chen JY, Schafer G, Horninger W, Lee C, Rubin MA, Klocker H, Demichelis F (2010) Genetic variation of genes involved in dihydrotestosterone metabolism and the risk of prostate cancer. Cancer Epidemiol Biomarkers Prev 19:229–239

    Article  PubMed  CAS  Google Scholar 

  41. Olsson M, Lindstrom S, Haggkvist B, Adami HO, Balter K, Stattin P, Ask B, Rane A, Ekstrom L, Gronberg H (2008) The UGT2B17 gene deletion is not associated with prostate cancer risk. Prostate 68:571–575

    Article  PubMed  CAS  Google Scholar 

  42. de Almagro MC, Selga E, Thibaut R, Porte C, Noe V, Ciudad CJ (2011) UDP-glucuronosyltransferase 1A6 overexpression in breast cancer cells resistant to methotrexate. Biochem Pharmacol 81:60–70

    Article  PubMed  Google Scholar 

  43. Justenhoven C, Winter S, Dunnebier T, Hamann U, Baisch C, Rabstein S, Spickenheuer A, Harth V, Pesch B, Bruning T, Ko YD, Brauch H (2010) Combined UGT1A1 and UGT1A6 genotypes together with a stressful life event increase breast cancer risk. Breast Cancer Res Treat 124:289–292

    Article  PubMed  Google Scholar 

  44. Starlard-Davenport A, Lyn-Cook B, Radominska-Pandya A (2008) Identification of UDP-glucuronosyltransferase 1A10 in non-malignant and malignant human breast tissues. Steroids 73:611–620

    Article  PubMed  CAS  Google Scholar 

  45. Chouinard S, Pelletier G, Belanger A, Barbier O (2006) Isoform-specific regulation of uridine diphosphate-glucuronosyltransferase 2B enzymes in the human prostate: differential consequences for androgen and bioactive lipid inactivation. Endocrinology 147:5431–5442

    Article  PubMed  CAS  Google Scholar 

  46. Turgeon D, Carrier JS, Levesque E, Hum DW, Belanger A (2001) Relative enzymatic activity, protein stability, and tissue distribution of human steroid-metabolizing UGT2B subfamily members. Endocrinology 142:778–787

    Article  PubMed  CAS  Google Scholar 

  47. Hu DG, Mackenzie PI (2009) Estrogen receptor alpha, fos-related antigen-2, and c-Jun coordinately regulate human UDP glucuronosyltransferase 2B15 and 2B17 expression in response to 17beta-estradiol in MCF-7 cells. Mol Pharmacol 76:425–439

    Article  PubMed  CAS  Google Scholar 

  48. Blevins-Primeau AS, Sun D, Chen G, Sharma AK, Gallagher CJ, Amin S, Lazarus P (2009) Functional significance of UDP-glucuronosyltransferase variants in the metabolism of active tamoxifen metabolites. Cancer Res 69:1892–1900

    Article  PubMed  CAS  Google Scholar 

  49. Sun D, Chen G, Dellinger RW, Duncan K, Fang JL, Lazarus P (2006) Characterization of tamoxifen and 4-hydroxytamoxifen glucuronidation by human UGT1A4 variants. Breast Cancer Res 8:R50

    Article  PubMed  Google Scholar 

  50. Farnham PJ, Means AL (1990) Sequences downstream of the transcription initiation site modulate the activity of the murine dihydrofolate reductase promoter. Mol Cell Biol 10:1390–1398

    PubMed  CAS  Google Scholar 

  51. Eroglu A, Egin Y, Cam R, Akar N (2009) The 19-bp deletion of dihydrofolate reductase (DHFR), methylenetetrahydrofolate reductase (MTHFR) C677T, Factor V Leiden, prothrombin G20210A polymorphisms in cancer patients with and without thrombosis. Ann Hematol 88:73–76

    Article  PubMed  CAS  Google Scholar 

  52. Guo Q, Wu M, Lian P, Liao M, Xiao Z, Wang X, Shen S (2009) Synergistic effect of indomethacin and NGX6 on proliferation and invasion by human colorectal cancer cells through modulation of the Wnt/beta-catenin signaling pathway. Mol Cell Biochem 330:71–81

    Article  PubMed  CAS  Google Scholar 

  53. Peng SP, Li XL, Wang L, Ou-Yang J, Ma J, Wang LL, Liu HY, Zhou M, Tang YL, Li WS, Luo XM, Cao L, Tang K, Shen SR, Li GY (2006) The role of NGX6 and its deletion mutants in the proliferation, adhesion and migration of nasopharyngeal carcinoma 5–8F cells. Oncology 71:273–281

    Article  PubMed  CAS  Google Scholar 

  54. Wang L, Ma J, Li J, Li X, Zhang Q, Peng S, Peng C, Zhou M, Xiong W, Yang J, Zhou J, Fan S, Tan C, Yan Q, Shen S, Li G (2005) NGX6 gene inhibits cell proliferation and plays a negative role in EGFR pathway in nasopharyngeal carcinoma cells. J Cell Biochem 95:64–73

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by a dissertation grant (M.Sc. thesis of EEN) from Zahedan University of Medical Sciences. The authors thank all subjects who willingly participated in the study.

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Authors and Affiliations

  1. Cellular and Molecular Research Center, Zahedan University of Medical Sciences, Zahedan, Iran

    Ebrahim Eskandari-Nasab & Mohammad Hashemi

  2. Department of Clinical Biochemistry, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

    Ebrahim Eskandari-Nasab, Mohammad Hashemi, Hamzeh Rezaei & Aliakbar Fazaeli

  3. Department of Internal Medicine, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

    Mohammad Ali Mashhadi & Farshid Arbabi

  4. Department of Pathology, Tamineh Ejtemaei Hospital, Zahedan, Iran

    Simin Sargholzaei Moghaddam

  5. Department of Pathology, School of Medicine, Zahedan University of Medical Sciences, Zahedan, Iran

    Mahdi Jahantigh

  6. Genetic of Non Communicable Disease Research Center, Zahedan University of Medical Sciences, Zahedan, Iran

    Mohsen Taheri

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  1. Ebrahim Eskandari-Nasab
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Eskandari-Nasab, E., Hashemi, M., Rezaei, H. et al. Evaluation of UDP-glucuronosyltransferase 2B17 (UGT2B17) and dihydrofolate reductase (DHFR) genes deletion and the expression level of NGX6 mRNA in breast cancer. Mol Biol Rep 39, 10531–10539 (2012). https://doi.org/10.1007/s11033-012-1938-8

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