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通过丁醇萃取和核酸酶P1增强的32P后标记法分析香烟烟雾诱导的人淋巴细胞和粒细胞中的DNA加合物。

Analysis of cigarette-smoke-induced DNA adducts by butanol extraction and nuclease P1-enhanced 32P-postlabeling in human lymphocytes and granulocytes.

作者信息

Savela K, Hemminki K

机构信息

Institute of Occupational Health, Topeliuksenkatu, Helsinki, Finland.

出版信息

Environ Health Perspect. 1993 Oct;101 Suppl 3(Suppl 3):145-50. doi: 10.1289/ehp.93101s3145.

Abstract

In an earlier study, we analyzed the aromatic DNA adducts separated from lymphocytes and granulocytes of smokers and nonsmokers using the nuclease P1-enhanced 32P-postlabeling assay. Here we compare the butanol extraction and nuclease P1-enhanced procedure on the same kind of samples. The DNA adducts of 42 per 10(8) nucleotides from smokers' lymphocytes were statistically higher (p < 0.05) than those of 11 from nonsmokers', when analyzed by the nuclease P1 treatment, but not by the 1-butanol extraction. The radioactivity obtained from the DNA digests on the TLC plates was lower in butanol-treated DNA samples when compared to those of nuclease P1 digestion. Lymphocytes appear to be a suitable test tissue for determining aromatic carcinogen exposure when detecting smoking-related DNA adducts by the nuclease P1-enhanced 32P-postlabeling analysis.

摘要

在一项早期研究中,我们使用核酸酶P1增强的32P后标记分析法,分析了从吸烟者和非吸烟者的淋巴细胞和粒细胞中分离出的芳香族DNA加合物。在此,我们比较了对同一样本进行丁醇萃取和核酸酶P1增强程序的效果。在用核酸酶P1处理时,吸烟者淋巴细胞每10⁸个核苷酸中有42个DNA加合物,在统计学上高于非吸烟者的11个(p < 0.05),但用1-丁醇萃取时则不然。与核酸酶P1消化的DNA样本相比,丁醇处理的DNA样本在薄层层析板上从DNA消化物中获得的放射性较低。当通过核酸酶P1增强的32P后标记分析检测与吸烟相关的DNA加合物时,淋巴细胞似乎是确定芳香族致癌物暴露的合适测试组织。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/25eb/1521148/e0dad5a7c290/envhper00380-0146-a.jpg

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