Thiol-modulated mechanisms of the cytotoxicity of thimerosal and inhibition of DNA topoisomerase II alpha.
Thimerosal is an organic mercury compound that is widely used as a preservative in vaccines and other solution formulations. The use of thimerosal has caused concern about its ability to cause neurological abnormalities due to mercury accumulation during a normal schedule of childhood vaccinations. While the chemistry and the biological effects of methylmercury have been well-studied, those of thimerosal have not. Thimerosal reacted rapidly with cysteine, GSH, human serum albumin, and single-stranded DNA to form ethylmercury adducts that were detectable by mass spectrometry. These results indicated that thimerosal would be quickly metabolized in vivo because of its reactions with protein and nonprotein thiols. Thimerosal also potently inhibited the decatenation activity of DNA topoisomerase II alpha, likely through reaction with critical free cysteine thiol groups. Thimerosal, however, did not act as a topoisomerase II poison and the lack of cross-resistance with a K562 cell line with a decreased level of topoisomerase II alpha (K/VP.5 cells) suggested that inhibition of topoisomerase II alpha was not a significant mechanism for the inhibition of cell growth. Depletion of intracellular GSH with buthionine sulfoximine treatment greatly increased the K562 cell growth inhibitory effects of thimerosal, which showed that intracellular glutathione had a major role in protecting cells from thimerosal. Pretreatment of thimerosal with glutathione did not, however, change its K562 cell growth inhibitory effects, a result consistent with the rapid exchange of the ethylmercury adduct among various thiol-containing cellular reactants. Thimerosal-induced single and double strand breaks in K562 cells were consistent with a rapid induction of apoptosis. In conclusion, these studies have elucidated some of the chemistry and biological activities of the interaction of thimerosal with topoisomerase II alpha and protein and nonprotein thiols and with DNA.
Wu
Xing
X
Faculty of Pharmacy, University of Manitoba, 50 Sifton Road, Winnipeg, Manitoba, R3T 2N2, Canada.
Liang
Hong
H
O'Hara
Kimberley A
KA
Yalowich
Jack C
JC
Hasinoff
Brian B
BB
D000951:Antigens, Neoplasm; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis / Q000187:drug effects; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors; D005978:Glutathione / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D011310:Preservatives, Pharmaceutical / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013438:Sulfhydryl Compounds / Q000737:chemistry / Q000378:metabolism*; D013849:Thimerosal / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D059005:Topoisomerase II Inhibitors
D000951:Antigens, Neoplasm; D018736:DNA Adducts; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D011310:Preservatives, Pharmaceutical; D012709:Serum Albumin; D013438:Sulfhydryl Compounds; D059005:Topoisomerase II Inhibitors; D013849:Thimerosal; D004250:DNA Topoisomerases, Type II; D005978:Glutathione; D003545:Cysteine