Vodicka P, Stetina R, Kumar R, Plna K, Hemminki K
Laboratory of Genetic Ecotoxicology, Regional Institute of Hygiene of Central Bohemia, Prague, Czech Republic.
Carcinogenesis. 1996 Apr;17(4):801-8. doi: 10.1093/carcin/17.4.801.
The modified 32P-postlabeling method was used for the detection of N-7-(2-hydroxy-phenylethyl) guanine adducts in DNA and human embryonal lung (HEL) cells treated in vitro with styrene oxide (SO). The total recovery of 7-alkylguanine adducts of styrene oxide in DNA analysed by 32P-postlabeling assay was 4.1 +/- 0.6%. The disappearance of 7-alkyldeoxyguanosine monophosphate adducts from SO-modified DNA at 37 degrees C showed a half-life of 19 h. The levels of 7-alkylguanine DNA adducts and single-strand breaks (SSBs) in DNA were determined in HEL cells treated with SO for 3 and 18 h. In the 3 h treatment there was a concentration-dependent increase of both 7-alkylguanine adducts and SSBs in DNA (r = 0.98, P = 0.012 and r = 0.99, P = 0.003, respectively). We found a significant correlation between 7-alkylguanine DNA adducts and SSBs in DNA (r = 0.98, P = 0.011). In HEL cells approximately 3-fold higher levels of both 7-alkylguanine DNA adducts and SSB in DNA were found after the SO (100 microM) treatment for 3 h than after the treatment for 18 h. A significant concentration-dependent increase was found only for SSB ( r = 0.95, P = 0.024) in the 18 h treatment with SO. There was no significant correlation between 7-alkylguanine adducts and SSBs (r = 0.72, P = 0.15). Our data suggest relatively fast removal of both 7-alkylguanine adducts of SO and DNA SSBs.
采用改良的³²P后标记法检测体外经环氧苯乙烯(SO)处理的DNA和人胚肺(HEL)细胞中的N-7-(2-羟基苯乙基)鸟嘌呤加合物。通过³²P后标记分析法分析,DNA中环氧苯乙烯的7-烷基鸟嘌呤加合物的总回收率为4.1±0.6%。在37℃下,SO修饰的DNA中7-烷基脱氧鸟苷单磷酸加合物的消失呈现出19小时的半衰期。对用SO处理3小时和18小时的HEL细胞中DNA的7-烷基鸟嘌呤加合物水平和单链断裂(SSB)进行了测定。在3小时的处理中,DNA中的7-烷基鸟嘌呤加合物和SSB均呈浓度依赖性增加(r分别为0.98,P = 0.012和r = 0.99,P = 0.003)。我们发现DNA中的7-烷基鸟嘌呤加合物与SSB之间存在显著相关性(r = 0.98,P = 0.011)。在HEL细胞中,用100μM的SO处理3小时后,DNA中的7-烷基鸟嘌呤加合物和SSB水平比处理18小时后高约3倍。在用SO处理18小时时,仅发现SSB有显著的浓度依赖性增加(r = 0.95,P = 0.024)。7-烷基鸟嘌呤加合物与SSB之间无显著相关性(r = 0.72,P = 0.15)。我们的数据表明,SO的7-烷基鸟嘌呤加合物和DNA单链断裂均可较快去除。