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Lack of association between GSTT1 polymorphism and endogenous or benzo[a]pyrene-induced sister chromatid exchanges as analyzed in metaphase or G2-phase lymphocytes

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Abstract

The objectives of the present work are (1) to verify whether genetic polymorphism in the detoxification gene GSTT1 influences the endogenous sensitivity in terms of sister chromatid exchanges (SCEs)/cell in healthy donors and (2) to test whether in vitro exposure to B[a]P in terms of SCEs/cell can be associated with polymorphism of GSTT1 gene. The presence or absence of the homozygous deletion in GSTT1 gene was determined in peripheral blood cells using multiplex-PCR. For SCEs quantitation, the cytogenetic method used thus far is based on the analysis in metaphase chromosomes. Consequently, G2-arrested cells are not included in the analysis. To overcome this shortcoming of the conventional method, we applied here SCE analysis in G2-phase prematurely condensed chromosomes (G2-PCCs) induced by calyculin-A, using a modified fluorescence-plus-Giemsa staining protocol. Compared to metaphase, a statistically significant increase in the yield of SCEs was notified in the G2-phase analysis after 48 h exposure of peripheral blood lymphocytes to 0.01–1 mM B[a]P, in both GSTT1-positive and -null donors. Therefore, the analysis of SCEs in the G2-phase using calyculin-A induced PCC methodology was shown to be more sensitive compared to the analysis at the metaphase level. Nevertheless, the results obtained do not show an association between the GSTT1 polymorphism with increased endogenous and/or B[a]P-induced SCE-frequencies in peripheral blood lymphocyte chromosomes in vitro. These results highlight not only the effect of B[a]P on cell cycle kinetics but also they demonstrate that conventional cytogenetic analysis at metaphase underestimates the cytogenetic effects of chemicals that delay cell cycle progression in G2-phase.

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References

  1. Ketterer B, Coles B, Meyer DJ (1983) The role of glutathione in detoxication. Environ Health Perspect 49:59–69

    Article  PubMed  CAS  Google Scholar 

  2. Dougherty D, Garte S, Barchowsky A, Zmuda J, Taioli E (2008) NQO1, MPO, CYP2e1, GSTT1 and GSTM1 polymorphisms and biological effects of benzene exposure—a literature review. Toxicol Lett 182(1–3):7–17

    Article  PubMed  CAS  Google Scholar 

  3. Piacentini S, Polimanti R, Porreca F, Martinez-Labarga C, De Stefano GF, Fuciarelli M (2010) GSTT1 and GSTM1 gene polymorphisms in European and African populations. Mol Biol Rep. doi:10.1007/s11033-010-0221-0

  4. Rafiee L, Saadat I, Saadat M (2010) Glutathione S-transferase genetic polymorphisms (GSTM1, GSTT1 and GSTO2) in three Iranian populations. Mol Biol Rep 37(1):155–158

    Article  PubMed  CAS  Google Scholar 

  5. Garte S, Gaspari L, Alexandrie AK, Ambrosone C, Autrup H, Autrup JL, Baranova H, Bathum L, Benhamou S, Boffetta P, Bouchardy C, Breskvar K, Brockmoller J, Cascorbi I, Clapper ML, Coutelle C, Daly A, Dell’Omo M, Dolzan V, Dresler CM, Fryer A, Haugen A, Hein DW, Hildesheim A, Hirvonen A, Hsieh LL, Ingelman-Sundberg M, Kalina I, Kang D, Kihara M, Kiyohara C, Kremers P, Lazarus P, Le Marchand L, Lechner MC, van Lieshout EM, London S, Manni JJ, Maugard CM, Morita S, Nazar-Stewart V, Noda K, Oda Y, Parl FF, Pastorelli R, Persson I, Peters WH, Rannug A, Rebbeck T, Risch A, Roelandt L, Romkes M, Ryberg D, Salagovic J, Schoket B, Seidegard J, Shields PG, Sim E, Sinnet D, Strange RC, Stucker I, Sugimura H, To-Figueras J, Vineis P, Yu MC, Taioli E (2001) Metabolic gene polymorphism frequencies in control populations. Cancer Epidemiol Biomarkers Prev 10(12):1239–1248

    PubMed  CAS  Google Scholar 

  6. Kelsey KT, Spitz MR, Zuo ZF, Wiencke JK (1997) Polymorphisms in the glutathione S-transferase class mu and theta genes interact and increase susceptibility to lung cancer in minority populations (Texas, United States). Cancer Causes Control 8(4):554–559

    Article  PubMed  CAS  Google Scholar 

  7. Altinisik J, Balta ZB, Aydin G, Ulutin T, Buyru N (2010) Investigation of glutathione S-transferase M1 and T1 deletions in lung cancer. Mol Biol Rep 37(1):263–267

    Article  PubMed  CAS  Google Scholar 

  8. Abdel-Rahman SZ, Anwar WA, Abdel-Aal WE, Mostafa HM, Au WW (1998) GSTM1 and GSTT1 genes are potential risk modifiers for bladder cancer. Cancer Detect Prev 22(2):129–138

    Article  PubMed  CAS  Google Scholar 

  9. Thakur H, Gupta L, Sobti RC, Janmeja AK, Seth A, Singh SK (2010) Association of GSTM1 T1 genes with COPD and prostate cancer in north Indian population. Mol Biol Rep. doi:10.1007/s11033-010-0287-8

  10. Saadat M, Saadat I (2010) Serum testosterone in females exposed to natural sour gas with respect to polymorphisms of XRCC1, GSTM1, and GSTT1. Mol Biol Rep. doi:10.1007/s11033-010-0081-7

  11. Tang JJ, Wang MW, Jia EZ, Yan JJ, Wang QM, Zhu J, Yang ZJ, Lu X, Wang LS (2010) The common variant in the gstm1 and gstt1 genes is related to markers of oxidative stress and inflammation in patients with coronary artery disease: a case-only study. Mol Biol Rep 37(1):405–410

    Article  PubMed  CAS  Google Scholar 

  12. Saadat I, Saadat M (2010) Influence of genetic polymorphisms of glutathione S-transferase T1 (GSTT1) and M1 (GSTM1) on haematological parameters. Mol Biol Rep 37(1):249–253

    Article  PubMed  CAS  Google Scholar 

  13. Chen H, Sandler DP, Taylor JA, Shore DL, Liu E, Bloomfield CD, Bell DA (1996) Increased risk for myelodysplastic syndromes in individuals with glutathione transferase theta 1 (GSTT1) gene defect. Lancet 347(8997):295–297

    Article  PubMed  CAS  Google Scholar 

  14. Masoudi M, Saadat I, Omidvari S, Saadat M (2009) Genetic polymorphisms of GSTO2, GSTM1, and GSTT1 and risk of gastric cancer. Mol Biol Rep 36(4):781–784

    Article  PubMed  CAS  Google Scholar 

  15. Azarpira N, Nikeghbalian S, Geramizadeh B, Darai M (2010) Influence of glutathione S-transferase m1 and t1 polymorphisms with acute rejection in Iranian liver transplant recipients. Mol Biol Rep 37(1):21–25

    Article  PubMed  CAS  Google Scholar 

  16. Saadat M (2009) Serum levels of testosterone and gonadotrophins with respect to smoking status and genetic polymorphism of GSTT1. Mol Biol Rep 36(6):1353–1356

    Article  PubMed  CAS  Google Scholar 

  17. Norppa H, Bonassi S, Hansteen IL, Hagmar L, Stromberg U, Rossner P, Boffetta P, Lindholm C, Gundy S, Lazutka J, Cebulska-Wasilewska A, Fabianova E, Sram RJ, Knudsen LE, Barale R, Fucic A (2006) Chromosomal aberrations and SCEs as biomarkers of cancer risk. Mutat Res 600(1–2):37–45

    PubMed  CAS  Google Scholar 

  18. Degrassi F, De Salvia R, Fiore M, Ricordy R, Tanzarella C, Benigni R, Palitti F (1993) Sensitivity of lymphocytes from vulcanizers to the in vitro induction of sister chromatid exchanges. Environ Health Perspect 101(3):97–100

    Article  PubMed  CAS  Google Scholar 

  19. Natarajan AT, Palitti F (2008) DNA repair and chromosomal alterations. Mutat Res 657(1):3–7

    PubMed  CAS  Google Scholar 

  20. Norppa H, Hirvonen A, Jarventaus H, Uuskula M, Tasa G, Ojajarvi A, Sorsa M (1995) Role of gstt1 and gstm1 genotypes in determining individual sensitivity to sister chromatid exchange induction by diepoxybutane in cultured human lymphocytes. Carcinogenesis 16(6):1261–1264

    Article  PubMed  CAS  Google Scholar 

  21. Sram RJ, Rossner P, Peltonen K, Podrazilova K, Mrackova G, Demopoulos NA, Stephanou G, Vlachodimitropoulos D, Darroudi F, Tates AD (1998) Chromosomal aberrations, sister-chromatid exchanges, cells with high frequency of SCE, micronuclei and comet assay parameters in 1,3-butadiene-exposed workers. Mutat Res 419(1–3):145–154

    PubMed  CAS  Google Scholar 

  22. Wiencke JK, Pemble S, Ketterer B, Kelsey KT (1995) Gene deletion of glutathione S-transferase theta: correlation with induced genetic damage and potential role in endogenous mutagenesis. Cancer Epidemiol Biomarkers Prev 4(3):253–259

    PubMed  CAS  Google Scholar 

  23. Bernardini S, Hirvonen A, Jarventaus H, Norppa H (2002) Influence of GSTM1 and GSTT1 genotypes on sister chromatid exchange induction by styrene in cultured human lymphocytes. Carcinogenesis 23(5):893–897

    Article  PubMed  CAS  Google Scholar 

  24. Norppa H (2001) Genetic polymorphisms and chromosome damage. Int J Hyg Environ Health 204(1):31–38

    Article  PubMed  CAS  Google Scholar 

  25. Bernardini S, Hirvonen A, Jarventaus H, Norppa H (2001) Trans-stilbene oxide-induced sister chromatid exchange in cultured human lymphocytes: influence of GSTM1 and GSTT1 genotypes. Mutagenesis 16(3):277–281

    Article  PubMed  CAS  Google Scholar 

  26. Landi S, Hanley NM, Warren SH, Pegram RA, DeMarini DM (1999) Induction of genetic damage in human lymphocytes and mutations in Salmonella by trihalomethanes: role of red blood cells and GSTT1–1 polymorphism. Mutagenesis 14(5):479–482

    Article  PubMed  CAS  Google Scholar 

  27. Yong LC, Schulte PA, Wiencke JK, Boeniger MF, Connally LB, Walker JT, Whelan EA, Ward EM (2001) Hemoglobin adducts and sister chromatid exchanges in hospital workers exposed to ethylene oxide: effects of glutathione S-transferase T1 and M1 genotypes. Cancer Epidemiol Biomarkers Prev 10(5):539–550

    PubMed  CAS  Google Scholar 

  28. Xiong P, Bondy ML, Li D, Shen H, Wang LE, Singletary SE, Spitz MR, Wei Q (2001) Sensitivity to benzo(a)pyrene diol-epoxide associated with risk of breast cancer in young women and modulation by glutathione S-transferase polymorphisms: a case-control study. Cancer Res 61(23):8465–8469

    PubMed  CAS  Google Scholar 

  29. Rojas M, Cascorbi I, Alexandrov K, Kriek E, Auburtin G, Mayer L, Kopp-Schneider A, Roots I, Bartsch H (2000) Modulation of benzo[a]pyrene diolepoxide-DNA adduct levels in human white blood cells by CYP1a1, GSTM1 and GSTT1 polymorphism. Carcinogenesis 21(1):35–41

    Article  PubMed  CAS  Google Scholar 

  30. Gupta RC, Earley K, Sharma S (1988) Use of human peripheral blood lymphocytes to measure DNA binding capacity of chemical carcinogens. Proc Natl Acad Sci USA 85(10):3513–3517

    Article  PubMed  CAS  Google Scholar 

  31. Bolt HM, Thier R (2006) Relevance of the deletion polymorphisms of the glutathione S-transferases gstt1 and gstm1 in pharmacology and toxicology. Curr Drug Metab 7(6):613–628

    Article  PubMed  CAS  Google Scholar 

  32. Malik SI, Terzoudi GI, Pantelias GE (2004) SCE analysis in G2 lymphocyte prematurely condensed chromosomes after exposure to atrazine: the non-dose-dependent increase in homologous recombinational events does not support its genotoxic mode of action. Cytogenet Genome Res 104(1–4):315–319

    Article  PubMed  CAS  Google Scholar 

  33. Terzoudi GI, Malik SI, Pantelias GE, Margaritis K, Manola K, Makropoulos W (2003) A new cytogenetic approach for the evaluation of mutagenic potential of chemicals that induce cell cycle arrest in the G2 phase. Mutagenesis 18(6):539–543

    Article  PubMed  CAS  Google Scholar 

  34. Jeffy BD, Chen EJ, Gudas JM, Romagnolo DF (2000) Disruption of cell cycle kinetics by benzo[a]pyrene: inverse expression patterns of BRCA-1 and p53 in MCF-7 cells arrested in S and G2. Neoplasia 2(5):460–470

    Article  PubMed  CAS  Google Scholar 

  35. Perry P, Wolff S (1974) New Giemsa method for the differential staining of sister chromatids. Nature 251(5471):156–158

    Article  PubMed  CAS  Google Scholar 

  36. Carrano AV, Natarajan AT (1988) International commission for protection against environmental mutagens and carcinogens. ICPEMC publication no. 14. Considerations for population monitoring using cytogenetic techniques. Mutat Res 204(3):379–406

    Article  PubMed  CAS  Google Scholar 

  37. Stavropoulou C, Korakaki D, Rigana H, Voutsinas G, Polyzoi M, Georgakakos VN, Manola KN, Karageorgiou CE, Sambani C (2007) Glutathione-S-transferase T1 and M1 gene polymorphisms in Greek patients with multiple sclerosis: a pilot study. Eur J Neurol 14(5):572–574

    Article  PubMed  CAS  Google Scholar 

  38. Schroder KR, Wiebel FA, Reich S, Dannappel D, Bolt HM, Hallier E (1995) Glutathione-S-transferase (GST) theta polymorphism influences background SCE rate. Arch Toxicol 69(7):505–507

    Article  PubMed  CAS  Google Scholar 

  39. Laffon B, Pasaro E, Mendez J (2002) Evaluation of genotoxic effects in a group of workers exposed to low levels of styrene. Toxicology 171(2–3):175–186

    Article  PubMed  CAS  Google Scholar 

  40. Kelsey KT, Wiencke JK, Ward J, Bechtold W, Fajen J (1995) Sister-chromatid exchanges, glutathione S-transferase theta deletion and cytogenetic sensitivity to diepoxybutane in lymphocytes from butadiene monomer production workers. Mutat Res 335(3):267–273

    PubMed  CAS  Google Scholar 

  41. Onaran I, Guven G, Ozaydin A, Ulutin T (2001) The influence of GSTM1 null genotype on susceptibility to in vitro oxidative stress. Toxicology 157(3):195–205

    Article  PubMed  CAS  Google Scholar 

  42. Guven GS, Guven M, Onaran I, Tunckale A, Hacihanefioglu S, Ulutin T (2005) Enhanced sensitivity to oxidant-induced micronucleus frequency in elderly individuals is not associated with glutathione S-transferase M1 (GSTM1) null genotype in lymphocytes. Gerontology 51(1):29–33

    Article  PubMed  CAS  Google Scholar 

  43. Laczmanska I, Gil J, Karpinski P, Stembalska A, Trusewicz A, Pesz K, Ramsey D, Schlade-Bartusiak K, Blin N, Sasiadek MM (2007) Polymorphism in nucleotide excision repair gene XPC correlates with bleomycin-induced chromosomal aberrations. Environ Mol Mutagen 48(8):666–671

    Article  PubMed  CAS  Google Scholar 

  44. Zhang Y, Rohde LH, Wu H (2009) Involvement of nucleotide excision and mismatch repair mechanisms in double strand break repair. Curr Genomics 10(4):250–258

    Article  PubMed  CAS  Google Scholar 

  45. Stavropoulou C, Sambani C, Rigana H, Georgakakos VN, Voutsinas G, Manola KN, Pantelias GE, Makropoulos V (2008) Low frequency of the glutathione-S-transferase T1-null genotype in patients with primary myelodysplastic syndrome and 5q deletion. Leukemia 22(8):1643–1646

    Article  PubMed  CAS  Google Scholar 

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Hatzi, V.I., Terzoudi, G.I., Stavropoulou, C. et al. Lack of association between GSTT1 polymorphism and endogenous or benzo[a]pyrene-induced sister chromatid exchanges as analyzed in metaphase or G2-phase lymphocytes. Mol Biol Rep 38, 3959–3966 (2011). https://doi.org/10.1007/s11033-010-0513-4

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