Cell-based assays for identification of novel double-strand break-inducing agents
Cell-based assays for identification of novel double-strand break-inducing agents
复制标题
DOI：
10.1093/jnci/94.2.88
复制
发表时间：
2002-01-16
影响因子：
10.3
通讯作者：
Lamb, JR
中科院分区：
文献类型：
--
作者：
Dunstan, HM;Ludlow, C;Lamb, JR
背景：我们正在开发以细胞为基础的检测方法，以确定对具有确定突变的细胞具有选择性毒性的抗癌药物。作为测试，我们使用三阶段策略从国家癌症研究所的储存库中筛选对双链断裂修复缺陷酵母细胞具有选择性毒性的化合物。研究方法：通过使用基于酵母和脊椎动物细胞的体外测定进一步分析筛选中鉴定的化合物，以区分拓扑异构酶I和II毒物。结果：从85000多个化合物中筛选出126个对DNA双链断裂修复缺陷酵母菌有选择性毒性的化合物。这126种化合物中有87种与已知的拓扑异构酶毒物结构相关，39种不相关。28的39个特征，我们目前的数据为8个化合物。在这八个化合物中，我们鉴定了两个新的拓扑异构酶11毒物（NSC 327929和NSC 638432）在生化试验和细胞中与依托泊苷等效，（NSC 63599，NSC 65601，NSC 380271，NSC 651646，和NSC 668370），其在酵母中具有拓扑异构酶I依赖性毒性，在哺乳动物细胞中诱导DNA损伤和毒性，以及一种（NSC 610898），其直接结合DNA并诱导链断裂。结论：基于细胞的测定可用于鉴定对具有预定遗传背景的细胞具有选择性毒性的分子，所述遗传背景包括参与细胞周期及其检查点的基因中的突变，对于所述细胞周期及其检查点，目前没有选择性毒性化合物。
Background: We are developing cell-based assays to identify anticancer agents that are selectively toxic to cells with defined mutations. As a test, we used a three-stage strategy to screen compounds from the National Cancer Institute's repository for agents that are selectively toxic to double-strand break repair-deficient yeast cells. Methods: Compounds identified in the screen were further analyzed by use of yeast and vertebrate cell-based and in vitro assays to distinguish between topoisomerase I and II poisons. Results: Of the more than 85 000 compounds screened, 126 were selectively toxic to yeast deficient in DNA double-strand break repair. Eighty-seven of these 126 compounds were structurally related to known topoisomerase poisons, and 39 were not. Twenty-eight of the 39 were characterized, and we present data for eight of the compounds. Among these eight compounds, we identified two novel topoisomerase 11 poisons (NSC 327929 and NSC 638432) that were equipotent to etoposide in biochemical tests and in cells, five (NSC 63599, NSC 65601, NSC 380271, NSC 651646, and NSC 668370) with topoisomerase I-dependent toxicity in yeast that induced DNA damage and toxicity in mammalian cells, and one (NSC 610898) that directly bound to DNA and induced strand breaks. Conclusions: Cell-based assays can be used to identify molecules that are selectively toxic to cells with a predetermined genetic background, including mutations in genes involved in the cell cycle and its checkpoints, for which there are currently no selectively toxic compounds.