Rosenstein B S
Department of Radiation Medicine, Brown University, Providence, RI 02912.
Environ Mol Mutagen. 1989;13(1):50-3. doi: 10.1002/em.2850130106.
ICR 2A frog cells and two solar ultraviolet (UV)-sensitive cell lines, DRP 36 and DRP 153, were irradiated with 150 kJ/m2 of the UV radiation produced by a fluorescent sun lamp, the radiation from which was passed through a sheet of 48A Mylar (DuPont, Wilmington, DE) to eliminate wavelengths shorter than approximately 315 nm. The irradiated cultures were also exposed to photoreactivating light (PRL), resulting in the removal of most of the pyrimidine dimers induced by the sun lamp UV irradiation, and then incubated 0-4 hr. At the end of the incubations, the cells were subjected to the alkaline elution assay. In these elutions, the cell lysates were either treated with proteinase K (proK) to eliminate any DNA-protein crosslinks (DPC) that may be present in the cells, or left untreated with proK. For the ICR 2A cells, the level of apparent DNA single-strand breaks (ssb) detected in elutions using proK increased with the incubation time after irradiation and remained high. However, when the DNA was eluted without proK pretreatment, the number of ssb fell rapidly. In contrast, the levels of ssb decreased in the DRP 36 and DRP 153 cells regardless of the use of proK in the elutions. Hence, this differential response in ssb induction may be indicative of a system involved with recovery following irradiation with solar UV wavelengths.
用荧光太阳灯产生的150 kJ/m²紫外线辐射照射ICR 2A青蛙细胞以及两种对太阳紫外线(UV)敏感的细胞系DRP 36和DRP 153,该辐射通过一片48A聚酯薄膜(杜邦公司,特拉华州威尔明顿)以滤除波长小于约315 nm的光。将受辐照的培养物也暴露于光复活光(PRL)下,从而去除太阳灯紫外线照射诱导产生的大部分嘧啶二聚体,然后孵育0至4小时。孵育结束时,对细胞进行碱性洗脱测定。在这些洗脱过程中,细胞裂解物要么用蛋白酶K(proK)处理以消除细胞中可能存在的任何DNA-蛋白质交联(DPC),要么不进行proK处理。对于ICR 2A细胞,使用proK进行洗脱时检测到的表观DNA单链断裂(ssb)水平随辐照后的孵育时间增加而升高并保持在较高水平。然而,当DNA洗脱前未用proK预处理时,ssb的数量迅速下降。相比之下,无论洗脱过程中是否使用proK,DRP 36和DRP 153细胞中的ssb水平均下降。因此,ssb诱导的这种差异反应可能表明存在一个与太阳紫外线波长辐照后恢复相关的系统。