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用32P后标记法检测对苯二酚和对苯醌处理的HL-60细胞中的DNA加合物。

Detection of DNA adducts in HL-60 cells treated with hydroquinone and p-benzoquinone by 32P-postlabeling.

作者信息

Levay G, Pongracz K, Bodell W J

机构信息

Department of Neurological Surgery, School of Medicine, University of California, San Francisco 94143.

出版信息

Carcinogenesis. 1991 Jul;12(7):1181-6. doi: 10.1093/carcin/12.7.1181.

Abstract

We have examined DNA adduct formation and cytotoxicity in HL-60 cells treated with either hydroquinone (HQ) or p-benzoquinone (p-BQ). Treatment of HL-60 cells with either HQ or p-BQ produced the same DNA adduct. The DNA adduct level varied from 0.05 to 10 adducts per 10(7) nucleotides as a function of treatment time and concentration for both compounds. To achieve the same DNA adduct level required higher concentrations and longer treatment times with HQ compared to p-BQ. p-BQ was also more cytotoxic to HL-60 cells than HQ. Reaction of calf thymus DNA with a p-BQ/HQ mixture produced five adducts as detected by 32P-postlabeling. Two isomers of (hydroxy)-1,N2-benzetheno-2'- deoxyguanosine-3'-phosphate were isolated from the reaction of 2'-deoxyguanosine-3'-phosphate with a p-BQ/HQ mixture and one of the isomers was identified as adduct no. 1 of the DNA reaction. The DNA adduct formed in HL-60 cells treated with HQ or p-BQ did not correspond to any of the principal adducts formed in DNA reacted with p-BQ/HQ. This result suggests that cellular mechanisms modify DNA adduct formation by HQ and p-BQ.

摘要

我们检测了用对苯二酚(HQ)或对苯醌(p-BQ)处理的HL-60细胞中的DNA加合物形成和细胞毒性。用HQ或p-BQ处理HL-60细胞产生了相同的DNA加合物。DNA加合物水平随两种化合物的处理时间和浓度而变化,范围为每10⁷个核苷酸0.05至10个加合物。与p-BQ相比,HQ达到相同的DNA加合物水平需要更高的浓度和更长的处理时间。p-BQ对HL-60细胞的细胞毒性也比HQ更大。通过³²P后标记检测,小牛胸腺DNA与p-BQ/HQ混合物反应产生了五种加合物。从2'-脱氧鸟苷-3'-磷酸与p-BQ/HQ混合物的反应中分离出两种(羟基)-1,N²-苯并乙烯基-2'-脱氧鸟苷-3'-磷酸异构体,其中一种异构体被鉴定为DNA反应的加合物编号1。用HQ或p-BQ处理的HL-60细胞中形成的DNA加合物与与p-BQ/HQ反应的DNA中形成的任何主要加合物均不对应。这一结果表明细胞机制改变了HQ和p-BQ导致的DNA加合物形成。

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