Measurement of unscheduled DNA synthesis in primary cultures of adult mouse epidermal keratinocytes.
Measurement of unscheduled DNA synthesis in primary cultures of adult mouse epidermal keratinocytes.
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成年小鼠表皮角质形成细胞原代培养物中非计划 DNA 合成的测量。
DOI：
10.1093/carcin/9.7.1197
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发表时间：
1988
期刊：
影响因子：
4.7
通讯作者：
DiGiovanni,J
中科院分区：
文献类型：
--
作者：
Sawyer,TW;Gill,RD;Smith-Oliver,T;Butterworth,BE;DiGiovanni,J
皮肤是许多环境致癌物的主要靶组织。为了提供一种工具来研究 NA 修复过程在该组织中与 DNA 损伤和癌变相关的作用，我们开发了一种测定方法，使用来自 SENCAR 小鼠的成年小鼠表皮角质形成细胞 (MEK) 培养物来测量化学诱导的 DNA 修复作为计划外 DNA 合成 (UDS)。制备初级 MEK，并在测试化学品和 10μCi /ml [3H]胸苷存在下孵育 24 小时。 UDS 通过放射自显影定量为每个核的净颗粒数。该测定检测了直接作用剂N-甲基-N'-硝基-N-亚硝基胍、N-甲基亚硝基脲、甲磺酸甲酯、甲磺酸乙酯和(±)-7α-8 α-二羟基-9α,10α-环氧-7,8,9,10-四氢苯并[a]芘和间接作用致癌物引起的DNA损伤。 7,12-二甲基苯并[a]蒽、苯并[a]芘、阿霉素和4-硝基喹啉-1-氧化物。所使用的生长条件允许表皮细胞保留体内小鼠表皮致癌物激活的组织特异性，因为2-乙酰氨基芴在该测定中是无活性的。该检测系统可用于确定皮肤的基因毒性和潜在致癌物质，以及涉及该组织中 DNA 修复和化学致癌作用的机制研究
Skin is a major target tissue for many environmental carcinogens. In order to provide a tool to study the role of NA-repair processes in relation to DNA damage and carcinogenesis in this tissue, we have developed an assay that measures chemically induced DNA repair as unscheduled DNA synthesis (UDS) using cultures of adult mouse epidermal keratinocytes (MEKs) from SENCAR mice. Primary MEKs were prepared and incubated for 24 h in the presence of the test chemical and 10μCi /ml [3H]thymidine. UDS was quantitated autoradiographically as net grains per nucleus. This assay detected DNA damage caused by the direct-acting agentsN-methyl-N′-nitro-N-nitrosoguanidine,N-methylnitrosourea, methyl methanesulfonate, ethyl methanesulfonate and (±)-7α-8 α-dihydroxy-9α,10α-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene and the indirect-acting carcinogens 7,12-dimethylbenz[a]anthracene, benzo[a]pyrene, adriamycin and 4-nitroquinoline-l-oxide. The growth conditions used allowed the epidermal cells to retain the tissue specificity of carcinogen activation of mouse epidermis in vivo in that 2-acetylaminofluorene was inactive in this assay. This assay system should be useful for determining genotoxic and potential carcinogenic agents for skin as well as for mechanistic studies involving DNA repair and chemical carcinogenesis in this tissue