Lack of association between GSTT1 polymorphism and endogenous or benzo[a]pyrene-induced sister chromatid exchanges as analyzed in metaphase or G2-phase lymphocytes.
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.
Health Physics Laboratory, Institute of Radioisotopes and Radiodiagnostic Products, National Center for Scientific Research, NCSR Demokritos, Terma Patriarhou Gregoriou and Neapoleos, Ag. Paraskevi Attikis, 153 10, Athens, Greece. vh@ipta.demokritos.gr
V I
VI
Hatzi
G I
GI
Terzoudi
C
C
Stavropoulou
S I
SI
Malik
V
V
Makropoulos
G E
GE
Pantelias
D001564:Benzo(a)pyrene / Q000494:pharmacology*; D016195:G2 Phase / Q000187:drug effects*; D005838:Genotype; D005982:Glutathione Transferase / Q000235:genetics*; D006801:Humans; D008214:Lymphocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D008677:Metaphase / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012854:Sister Chromatid Exchange / Q000187:drug effects*; D014019:Tissue Donors
D001564:Benzo(a)pyrene; C413545:glutathione S-transferase T1; D005982:Glutathione Transferase