BULKY DNA-ADDUCT FORMATION INDUCED BY NI(II) INVITRO AND INVIVO AS ASSAYED BY P-32 POSTLABELING

被引:33
作者
CHANG, J
WATSON, WP
RANDERATH, E
RANDERATH, K
机构
[1] BAYLOR COLL MED,DEPT PHARMACOL,DIV TOXICOL,HOUSTON,TX 77030
[2] SHELL RES LTD,SITTINGBOURNE RES CTR,SITTINGBOURNE ME9 8AG,KENT,ENGLAND
来源
MUTATION RESEARCH | 1993年 / 291卷 / 02期
关键词
DNA ADDUCTS; NICKEL CARCINOGENESIS; OXYGEN FREE RADICALS; FENTON REACTION; P-32-POSTLABELING;
D O I
10.1016/0165-1161(93)90154-R
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Various small oxidation products (e.g. 8-hydroxydeoxyguanosine) can be induced in DNA by nickel compounds. In this study, the P-32-postlabeling assay was applied to determine whether Ni(II) compounds are able to induce bulky DNA-adduct formation in vitro and in vivo. In vitro studies detected two major and several minor adducts in DNA incubated with NiCl2 and H2O2 at 37-degrees-C for 1 h. Formation of the two major adducts increased with incubation time (0-24 h) and NiCl2 concentration (0-800 muM). Adduct levels were greatly reduced by hydroxyl free-radical scavengers, i.e. 0.4 M sodium formate or 0.05 M p-nitrosodimethylaniline, and by a singlet oxygen scavenger, 0.05 M sodium azide. The in vitro effects of NiCl2 on DNA were significantly enhanced by (1) addition of 3 mM ascorbic acid, (2) replacement of H2O with D2O in the reaction, and (3) prior denaturation of DNA. Adduct formation presumably involved a Fenton-type reaction, in which DNA crosslinks may arise by reaction with hydroxyl free radicals and singlet oxygen. For in vivo studies, male 6-8 wk old B6C3F1 mice were used. In untreated mice, several 1-compounds (putative indigenous DNA modifications that increase with age) were detected in liver, kidney, and lung. Two of these (spots 1 and 2) were chromatographically identical to the two major spots induced by Ni(II) in vitro. The intensities of spots 1 and 2 in kidney and of some other spots in liver and lung were increased 1 and 2 h after i.p. injection with a single dose of 170 mumoles/kg NiAc2. The effects of NiAc, were reduced or undetectable in the three tissues 24 h after treatment. These observations indicate the capacity of Ni(II) to induce and modulate bulky DNA modifications both in vitro and in vivo.
引用
收藏
页码:147 / 159
页数:13
相关论文
共 59 条
[51]   ASSOCIATION BETWEEN ERYTHROCYTOSIS AND RENAL CANCERS IN RATS FOLLOWING INTRARENAL INJECTION OF NICKEL COMPOUNDS [J].
SUNDERMAN, FW ;
MCCULLY, KS ;
HOPFER, SM .
CARCINOGENESIS, 1984, 5 (11) :1511-1517
[52]  
SUNDERMAN FW, 1983, ORGAN SPECIES SPECIF, P107
[53]  
SUNDERMAN FW, 1984, IARC SCI PUBL, V53, P127
[54]  
WANG XW, 1988, TOXICOLOGIST, V8, P15
[55]   POST-RADIOLABELING FOR DETECTING DNA DAMAGE [J].
WATSON, WP .
MUTAGENESIS, 1987, 2 (05) :319-331
[56]  
Wills E.D., 1985, OXIDATIVE STRESS, P197
[58]   HIGH-RESOLUTION TLC MAPPING AND CHARACTERIZATION OF P-32 POSTLABELED MONOPHOSPHATE 7,12-DIMETHYLBENZ[A]ANTHRACENE DNA ADDUCTS [J].
YANG, PF ;
RANDERATH, K .
CARCINOGENESIS, 1990, 11 (01) :159-164
[59]  
1986, EPA600883012FF