Seregina T B, Kuz'min I A, Zhestianikov V D
Tsitologiia. 1996;38(1):57-65.
A method of sedimentation in alkaline sucrose gradients was used to study repair of gamma-induced DNA single strand breaks (SSB) and DNA degradation in HeLa cells treated with novobiocin (Nb), an inhibitor of topoisomerase II. After irradiation in a dose of 150 Gr, Nb in a concentration of 1 mM does not affect the effectivity of SSB repair and DNA molecular mass in the irradiated cells treated with Nb for 60-180 min before irradiation. Besides, it does not lead to additional DNA degradation in cells treated with Nb for 60-180 min before gamma-rays, as well as following a postirradiation incubation during 60-180 min. Nb in a concentration of 4 mM, much exceeding the Nb concentration when DNA synthesis and cell transit through the cycle are inhibited, causes the following changes. It does not affect DNA molecular mass in non-irradiated cells, inhibits repair of DNA SSB, causes partial DNA degradation, if cells are treated for 60-180 min before gamma-rays and during the following postirradiation incubation (60-180 min). Taking into account the Nb-mediated DNA degradation, the inhibition of DNA repair by Nb appears not significant. Since in a concentration, which inhibits topoisomerase II, Nb does not affect repair of gamma-induced DNA SSB, one may assume the lack of involvement of topoisomerase II into repair of these DNA lesions. Inhibition of DNA repair by 4 mM Nb may result from its effect on the number of proteins, including reparative DNA polymerase, rather than from a selective effect on topoisomerase II.
采用碱性蔗糖梯度沉降法研究了用拓扑异构酶II抑制剂新生霉素(Nb)处理的HeLa细胞中γ射线诱导的DNA单链断裂(SSB)的修复和DNA降解情况。在150格雷剂量照射后,浓度为1 mM的Nb对在照射前用Nb处理60 - 180分钟的照射细胞中SSB修复效率和DNA分子量没有影响。此外,它不会导致在γ射线照射前用Nb处理60 - 180分钟的细胞以及照射后60 - 180分钟孵育期间出现额外的DNA降解。浓度为4 mM的Nb大大超过了抑制DNA合成和细胞周期进程时的Nb浓度,会引起以下变化。它对未照射细胞的DNA分子量没有影响,抑制DNA SSB的修复,如果在γ射线照射前和照射后孵育期间(60 - 180分钟)用细胞处理60 - 180分钟,会导致部分DNA降解。考虑到Nb介导的DNA降解,Nb对DNA修复的抑制作用似乎不显著。由于在抑制拓扑异构酶II的浓度下,Nb不影响γ射线诱导的DNA SSB的修复,因此可以推测拓扑异构酶II不参与这些DNA损伤的修复。4 mM Nb对DNA修复的抑制可能是由于其对包括修复性DNA聚合酶在内的多种蛋白质数量的影响,而不是对拓扑异构酶II的选择性作用。