Oesch F, Weiss C M, Klein S
Institute of Toxicology, University of Mainz, Germany.
Arch Toxicol. 1994;68(6):358-63. doi: 10.1007/s002040050082.
Oligonucleotide duplexes of a defined sequence containing one 1,N6-ethenoadenosine (EA) were synthesized and used as substrates to study the repair of this DNA lesion in cell homogenates of peripheral mononuclear blood cells of 39 male and female workers, exposed to vinyl chloride. These data were compared to data from 39 employees of the same company working in other production plants and to data from a control group of 39 persons, living in an area without vinyl chloride production. After incubation of the 5'- and 3'-labeled oligonucleotide duplex with cell homogenate, a specific nicking activity, releasing the deoxyribosyl phosphate originally carrying the EA, was found. This activity was used to determine the individual and collective repair activities for ethenoadenine. The exposed group showed a mean of 158.5 +/- 39.9 (SD) fmol product fragment and did not differ significantly from the mean value of the two control groups with 156.5 +/- 42.9 fmol and 161.2 +/- 53.6 fmol, respectively. Large interindividual variations were found, ranging from 4.9-fold in the exposed to 8.2- and 7.2-fold in the control groups. The development of an assay for ethenoadenine repair is significant for understanding the role of EA repair in eukaryotic cells.
合成了含有一个1,N6-乙烯腺嘌呤(EA)的特定序列的寡核苷酸双链体,并将其用作底物,以研究39名接触氯乙烯的男性和女性工人外周血单核细胞匀浆中这种DNA损伤的修复情况。将这些数据与同一家公司在其他生产车间工作的39名员工的数据以及来自生活在无氯乙烯生产地区的39名对照组人员的数据进行比较。在用细胞匀浆孵育5'-和3'-标记的寡核苷酸双链体后,发现了一种特异性切口活性,释放出原本携带EA的脱氧核糖磷酸。该活性用于确定乙烯腺嘌呤的个体和集体修复活性。暴露组的平均产物片段为158.5±39.9(标准差)fmol,与两个对照组的平均值分别为156.5±42.9 fmol和161.2±53.6 fmol相比,无显著差异。发现个体间差异很大,暴露组的差异为4.9倍,对照组为8.2倍和7.2倍。开发一种乙烯腺嘌呤修复检测方法对于理解EA修复在真核细胞中的作用具有重要意义。