DETECTION AND MEASUREMENT OF HUMAN DNA ADDUCTS

人类 DNA 加合物的检测和测量

基本信息

批准号：
3548870
负责人：
KURT RANDERATH
金额：
$ 16.36万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1986
资助国家：
美国
起止时间：
1986-08-01 至 1990-07-31
项目状态：
已结题

项目摘要

A 32P-Postlabeling method developed in the applicant's laboratory will be applied to study potential covalent DNA lesions in human white blood cells and tissues exposed to occupational or environmental mixtures of genotoxic compounds. The basic procedure entails the enzymatic digestion of the DNA preparation to be tested for the presence of adducts to deoxyribonucleoside 3'-monophosphates, conversion of these mononucleotides to 5'-32P-labeled deoxyribonucleoside 3',5'-bisphosphates, resolution of the 32P-labeled nucleotides by thin-layer chromatography, and autoradiography. 32P-Labeled adduct fractions are detected as extra spots on X-ray film and are quantitated by scintillation (Cerenkov) counting. Characteristic "fingerprints" are obtained for DNA adducts formed in vivo from authentic carcinogens or mixtures of genotoxic compounds such as cigarette smoke. The 32P-postlabeling assay will be suitable specifically for the analysis of traces of adducts in microgram amounts of human DNA. For the characterization of adducts observed in human DNA, co-chromatography and stability studies will be conducted with adducts obtained from DNA of experimental animals treated with crude environmental/occupational mixtures or authentic carcinogens known to occur in the exposure of interest. Cigarette smoking will be considered as a confounding variable on the basis of the detection of specific smoking-associated DNA adducts. The project will focus on occupational exposures to polycyclic aromatic hydrocarbons, aromatic amines/amides, azo compounds, dyestuffs, formaldehyde, and styrene. In addition, it is proposed to study whether the exposure to emissions of wood-burning stoves and the natural process of aging give rise to adducts in human DNA.

申请人实验室开发的 32P-Postlabeling 方法将用于研究人类白细胞中潜在的共价 DNA 损伤以及暴露于职业或环境遗传毒性混合物的组织化合物。基本程序包括 DNA 的酶消化待测试脱氧核糖核苷加合物存在的制剂 3'-单磷酸，将这些单核苷酸转化为 5'-32P 标记脱氧核糖核苷 3',5'-二磷酸，32P 标记的分辨率通过薄层色谱法和放射自显影法检测核苷酸。 32P-标记加合物部分被检测为 X 射线胶片上的额外斑点，并被通过闪烁（切伦科夫）计数进行定量。特征 “指纹”是从真实的体内形成的DNA加合物获得的致癌物或遗传毒性化合物的混合物，例如香烟烟雾。 32P 标记后测定特别适合分析微克量的人类 DNA 中的加合物痕迹。为了表征在人类 DNA 中观察到的加合物，将使用加合物进行共色谱和稳定性研究取自经粗处理的实验动物的DNA 已知发生的环境/职业混合物或真正的致癌物质在利益的曝光中。吸烟将被视为基于特定检测的混杂变量与吸烟相关的 DNA 加合物。该项目将重点关注多环芳烃的职业暴露碳氢化合物、芳香胺/酰胺、偶氮化合物、染料、甲醛和苯乙烯。此外，建议研究是否暴露于燃木炉的排放物和自然过程衰老会导致人类 DNA 中产生加合物。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Aromatic DNA adducts in white blood cells of coke workers.

焦炭工人白细胞中的芳香族 DNA 加合物。

DOI：
10.1007/bf00379065
发表时间：
1990
期刊：
International archives of occupational and environmental health
影响因子：
3
作者：
Hemminki,K;Grzybowska,E;Chorazy,M;Twardowska-Saucha,K;Sroczynski,JW;Putman,KL;Randerath,K;Phillips,DH;Hewer,A
通讯作者：
Hewer,A

Tissue distribution of covalent DNA damage in mice treated dermally with cigarette 'tar': preference for lung and heart DNA.

用香烟“焦油”进行皮肤处理的小鼠中共价 DNA 损伤的组织分布：对肺和心脏 DNA 的偏好。