Hsia M T, Kreamer B L, Dolara P
Mutat Res. 1983 Nov;122(2):177-85. doi: 10.1016/0165-7992(83)90057-x.
The filter elution method used for the detection of DNA strand breaks has been modified to quantitate chemically induced DNA repair which is measured as unscheduled DNA synthesis (UDS) in suspension of freshly isolated rat hepatocytes. Our method is based on DNA purification by retention on polyvinyl chloride filters, and is capable of handling a large number of samples simultaneously. By using the present assay system, positive dose-dependent UDS data was obtained on the following carcinogens: aflatoxin B1, 2-acetylaminofluorene, 4-aminobiphenyl, 2-aminofluorene, methyl methanesulfonate, N-methyl-N'-nitro-N-nitrosoguanidine, and 4-nitroquinoline-1-oxide. In contrast, non-carcinogenic biphenyl, fluorene, and sodium ascorbate did not elicit any detectable levels of UDS at all concentrations tested. Thus, UDS as measured by the present filter retention method may serve as an efficient and reliable means of screening chemical mutagens/carcinogens.
用于检测DNA链断裂的滤膜洗脱法已得到改进，以定量化学诱导的DNA修复，该修复在新鲜分离的大鼠肝细胞悬液中通过非预定DNA合成（UDS）来测定。我们的方法基于通过保留在聚氯乙烯滤膜上来纯化DNA，并且能够同时处理大量样品。通过使用本检测系统，在以下致癌物上获得了阳性剂量依赖性UDS数据：黄曲霉毒素B1、2-乙酰氨基芴、4-氨基联苯、2-氨基芴、甲磺酸甲酯、N-甲基-N'-硝基-N-亚硝基胍和4-硝基喹啉-1-氧化物。相比之下，非致癌性的联苯、芴和抗坏血酸钠在所有测试浓度下均未引发任何可检测水平的UDS。因此，通过本滤膜保留法测定的UDS可作为筛选化学诱变剂/致癌物的有效且可靠的手段。
