A 32P-postlabeling method for the detection of adducts in the DNA of human fibroblasts exposed to sulfur mustard

Chemico-biological Interactions
T Q NiuD B Ludlum

Abstract

Since the toxicities of sulfur mustard are attributed to DNA alkylation, levels of DNA modification in exposed cells should correlate with the intensity of exposure. We have found that 32P-postlabeling can be used successfully to detect the major adduct, 7-hydroxyethylthio- ethyldeoxyguanosine 5'-phosphate (HETEpdG), that is formed in DNA by sulfur mustard. This method has been used to establish a correlation between exposure and adduct formation in human fibroblasts grown in cell culture and exposed to sulfur mustard concentrations between 2.5 and 15 microM. DNA was recovered from these cells using a salt precipitation method to remove proteins and was found to have an HETEpdG content which increased linearly with SM concentration. This relationship shows that one HETEpdG per 10(6) nucleotides is produced at a SM concentration of 2.3 microM. Growth of fibroblast cells, assayed by trypan-blue exclusion, is somewhat inhibited by 2 microM SM, indicating that 32P-postlabeling has the requisite sensitivity to detect adducts at levels of SM that are minimally toxic.

References

Feb 11, 1988·Nucleic Acids Research·S A MillerH F Polesky
Jul 19, 1994·Proceedings of the National Academy of Sciences of the United States of America·D YuD B Ludlum
May 1, 1994·Chemical Research in Toxicology·G P van der SchansR A Baan
Apr 1, 1994·Chemico-biological Interactions·D B LudlumD Yu

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Citations

Feb 22, 2001·Biochemical and Biophysical Research Communications·A H FangR C Gupta
Nov 14, 2003·Toxicology and Applied Pharmacology·Lei MiThomas W Sawyer
Oct 29, 2005·American Journal of Respiratory and Critical Care Medicine·Mostafa GhaneiJafar Aslani
May 5, 1999·Chemico-biological Interactions·X M ChenD R Phillips
Nov 21, 2001·Chemico-biological Interactions·S ShahinP J Gray
Aug 4, 2016·Journal of Receptor and Signal Transduction Research·Mohammad Reza NouraniAbbas Ali Imani Fooladi
Aug 9, 2005·Toxicology·Kai Kehe, Ladislaus Szinicz
May 27, 2014·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yajiao ZhangJianwei Xie
Dec 1, 2004·Nutrition Research Reviews·Yves Ingenbleek, Vernon R Young
Jun 25, 2019·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·Stephen C Mitchell, Glyn B Steventon

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