Journal Article

7-Alkylguanine adducts of styrene oxide determined by <sup>32</sup>P-postlabelling in DNA and human embryonal lung fibroblasts (HEL)

Pavel Vodicka, Rudolf Stetina, Rajiv Kumar, Kamila Plna and Kari Hemminki

in Carcinogenesis

Volume 17, issue 4, pages 801-808
Published in print April 1996 | ISSN: 0143-3334
Published online April 1996 | e-ISSN: 1460-2180 | DOI: https://dx.doi.org/10.1093/carcin/17.4.801
7-Alkylguanine adducts of styrene oxide determined by
                    32P-postlabelling in DNA and human embryonal lung fibroblasts
                    (HEL)

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The modified 32P-postlabelling method was used for the detection of N-7-(2-hydroxy-phenylethyl) guanine adducts in DNA and human embryonal lung (HEL) cells treated in vitro with styrene oxide (SO). The total recovery of 7-alkylguanine adducts of styrene oxide in DNA analysed by 32P-postlabelling assay was 4.1±0.6%. The disappearance of 7-alkyldeoxyguanosine monophosphate adducts from SO-modified DNA at 37°C showed a half-life of 19 h. The levels of 7-alkylguanine DNA adducts and single-strand breaks (SSBs) in DNA were determined in HEL cells treated with SO for 3 and 18 h. In the 3 h treatment there was a concentration-dependent increase of both 7-alkylguanine adducts and SSBs in DNA (r = 0.98, P = 0.012 and r = 0.99, P = 0.003, respectively). We found a significant correlation between 7-alkylguanine DNA adducts and SSBs in DNA (r = 0.98, P = 0.011). In HEL cells ∼3-fold higher levels of both 7-alkylguanine DNA adducts and SSBs in DNA were found after the SO (100 μM) treatment for 3 h than after the treatment for 18 h. A significant concentration-dependent increase was found only for SSB(r = 0.95, P = 0.024) in the 18 h treatment with SO. There was no significant correlation between 7-alkylguanine adducts and SSBs (r = 0.72, P = 0.15). Our data suggest relatively fast removal of both 7-alkylguanine adducts of SO and DNA SSBs.

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Subjects: Clinical Cytogenetics and Molecular Genetics

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