Induction of direct adducts, apurinic/apyrimidinic sites and oxidized bases in nuclear DNA of human HeLa S3 tumor cells by tetrachlorohydroquinone.
DNA damage induced by tetrachlorohydroquinone (Cl(4)HQ), the quinonoid metabolite of pentachlorophenol (PCP), was investigated in human HeLa S3 tumor cells. Formation of one major and two minor DNA adducts in cells treated with Cl(4)HQ (50-300 microM) was detected by (32)P-post-labeling assay and the adducts accumulated over the course of the experiment (0.5-2 h), with total adduct levels estimated to be 3-6 per 10(8) nucleotides. These adducts did not correspond to those derived from calf thymus DNA treated with tetrachloro-1,4-benzoquinone. Results from the apurinic/apyrimidinic (AP) sites assay indicated that the number of AP sites was 2-fold greater in cells exposed to Cl(4)HQ (300 microM) than the corresponding control. Further characterization of the AP sites confirmed that Cl(4)HQ induced predominantly (75%) putrescine-excisable AP sites in HeLa S3 cells. In parallel, the concentration of 8-hydroxy-2'-deoxyguanosine (8-HO-dG) in cells treated with Cl(4)HQ for 0.5 and 2 h was increased 2- and 5-fold, respectively, compared with the control. The extent of oxidative DNA damage induced by Cl(4)HQ was approximately two orders of magnitude greater than those of direct DNA adducts. Overall, it appears that reactive oxygen species mediate the parallel formation of AP sites and 8-HO-dG in HeLa S3 cells following treatment with Cl(4)HQ and that the contribution of depurination/depyrimidination of direct DNA adducts is relatively insignificant compared with the formation of oxidized AP sites. We conclude that putrescine-excisable AP sites represent a major type of ROS-mediated oxidative DNA damage in cellular DNA induced by Cl(4)HQ and may play a role in PCP-induced clastogenicity in mammalian cells.
Lin
P H
PH
Department of Environmental Sciences and Engineering, The University of North Carolina, Chapel Hill, NC 27599-7400, USA.
Nakamura
J
J
Yamaguchi
S
S
La
D K
DK
Upton
P B
PB
Swenberg
J A
JA
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002467:Cell Nucleus / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000187:drug effects*; D018736:DNA Adducts; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D006367:HeLa Cells; D006801:Humans; D006873:Hydroquinones; D008956:Models, Chemical; D010100:Oxygen / Q000378:metabolism*; D011687:Purines; D011700:Putrescine / Q000494:pharmacology; D011743:Pyrimidines; D017382:Reactive Oxygen Species; D013997:Time Factors
D018736:DNA Adducts; D006873:Hydroquinones; D011687:Purines; D011743:Pyrimidines; D017382:Reactive Oxygen Species; C044578:2,3,5,6-tetrachlorohydroquinone; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; D003849:Deoxyguanosine; D010100:Oxygen; D011700:Putrescine