Cortés F, Daza P, Piñero J, Escalza P
Department of Cell Biology, Faculty of Biology of Seville, Spain.
Environ Mol Mutagen. 1994;24(3):203-7. doi: 10.1002/em.2850240309.
An approach based on the synchronization of CHO cells after a first cell cycle incorporating a relatively low amount of bromodeoxyuridine (BrdUrd) into DNA, followed by mutagenic treatment and subsequent culture for second and third generations of BrdUrd incorporation for the scoring of sister chromatid exchanges (SCEs) per cell cycle in three-way differentially (TWD) stained chromosomes, has been used to investigate the possible cancellation of SCEs. Cancellation is expected to occur if two mutagen-induced SCEs occur at exactly the same site in subsequent rounds of replication. Lesions in DNA seem to persist and are able to induce SCE throughout two cell cycles after treatment with the three mutagens tested--mitomycin C (MMC), ethyl methanesulfonate (EMS) and ultraviolet (UV) light--though this latter agent was shown as only moderately persistent. Our results seem to indicate that SCEs induced by these mutagens do not take place at the same locus in successive cell generations, as assessed by a lack of SCE cancellation.
一种方法是,在第一个细胞周期后使中国仓鼠卵巢(CHO)细胞同步化,让相对少量的溴脱氧尿苷(BrdUrd)掺入DNA,接着进行诱变处理,随后培养第二代和第三代掺入BrdUrd的细胞,用于在经三色差别(TWD)染色的染色体中对每个细胞周期的姐妹染色单体交换(SCE)进行评分,以此来研究SCE可能的消除情况。如果在随后的复制轮次中,两个诱变诱导的SCE恰好发生在同一位置,预计会发生消除。在用三种测试诱变剂——丝裂霉素C(MMC)、甲基磺酸乙酯(EMS)和紫外线(UV)——处理后,DNA损伤似乎会持续存在,并能够在两个细胞周期内诱导SCE,不过后一种诱变剂显示其持久性仅为中等程度。我们的结果似乎表明,通过缺乏SCE消除来评估,这些诱变剂诱导的SCE并非在连续细胞世代的同一基因座发生。