Influence of o-phenanthroline on DNA single-strand breaks, alkali-labile sites, glutathione reductase, and formation of chromium(V) in Chinese hamster V-79 cells treated with sodium chromate (VI).
The effect of the cell-permeable metal chelator o-phenanthroline (OP) on the formation of chromium(V), DNA breaks, alkali-labile sites, and enzyme inhibition of chromium(VI) was studied using Chinese hamster V-79 cells. Alkaline elution assays demonstrated that treatment with OP (100-500 microM) resulted in a dose-dependent decrease in cellular levels of either alkali-labile sites or the combination of alkali-labile sites plus DNA single-strand breaks caused by Na2CrO4. Cellular treatment with OP also attenuated the inhibition of glutathione reductase attributed to Na2CrO4. Under the same experimental conditions, the cellular uptake of chromate was not affected by OP. ESR studies revealed that cellular treatment with OP (100-500 microM) resulted in a concentration-dependent decrease in the level of chromium(V) intermediate in cells treated with Na2CrO4. Furthermore, OP inhibited the chromium(V) complex that formed during the reaction of Na2CrO4 with glutathione in vitro, resulting in the decrease of chromium(V)-mediated hydroxyl radical formation while neither the reduction of chromium(VI) nor the formation of the chromium(VI)-glutathione complex was affected, indicating that OP may react with chromium(V) but not with chromium(VI). These results suggest that the metal chelator OP decreases chromate-induced alkali-labile sites or the combination of alkali-labile sites plus DNA strand breaks, as well as chromium inhibition of glutathione reductase, possibly through its ability to directly decrease chromium(V) complex in cells.
Department of Medical Biochemistry, Kurume University School of Medicine, Japan.
M
M
Sugiyama
K
K
Tsuzuki
N
N
Haramaki
D000818:Animals; D002460:Cell Line; D002857:Chromium / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004249:DNA Damage; D004578:Electron Spin Resonance Spectroscopy; D005980:Glutathione Reductase / Q000378:metabolism*; D066298:In Vitro Techniques; D010084:Oxidation-Reduction; D010618:Phenanthrolines / Q000494:pharmacology*
D010618:Phenanthrolines; D002857:Chromium; D004247:DNA; D005980:Glutathione Reductase; C025205:1,10-phenanthroline