An image-based, high-throughput screening assay for molecules that induce excess DNA replication in human cancer cells.
An image-based, high-throughput screening assay for molecules that induce excess DNA replication in human cancer cells.
复制标题
一种基于图像的高通量筛选方法，用于检测在人类癌细胞中诱导过度 DNA 复制的分子。
DOI：
10.1158/1541-7786.mcr-10-0570
复制
发表时间：
2011
期刊：
影响因子：
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通讯作者：
DePamphilis,MelvinL
中科院分区：
文献类型：
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作者：
Zhu,Wenge;Lee,ChrissieY;Johnson,RonaldL;Wichterman,Jennifer;Huang,Ruili;DePamphilis,MelvinL
以前的研究表明，在正常组织来源的细胞不可能的条件下，可以在人类癌症来源的细胞中诱导DNA再复制。由于DNA再复制诱导细胞死亡，这种策略可以应用于发现潜在的抗癌疗法。因此，开发了适合于高通量筛选的成像测定，其测量完整细胞核中超过四个基因组当量的DNA复制并指示细胞增殖。通过在正常和肿瘤衍生细胞上筛选1，280种生物活性分子的文库来验证该测定法，其中证明其比目前用于检测过量DNA复制的方法更灵敏。该筛选鉴定了已知的过量DNA复制诱导剂，如微管动力学抑制剂，以及在正常细胞和癌细胞中诱导过量DNA复制的新型化合物。此外，鉴定了两种化合物，其在癌细胞中选择性地诱导过量DNA复制，并且一种化合物在癌细胞中选择性地诱导内循环。因此，该测定法提供了一种发现可用于研究基因组复制的调节和用于治疗癌症的化合物的新方法（Mol Cancer Res; 9（3）; 294-310）。©2011 AACR。
Previous studies have shown DNA re-replication can be induced in cells derived from human cancers under conditions in which it is not possible for cells derived from normal tissues. Because DNA re-replication induces cell death, this strategy could be applied to the discovery of potential anticancer therapeutics. Therefore, an imaging assay amenable to high-throughput screening was developed that measures DNA replication in excess of four genomic equivalents in the nuclei of intact cells and indexes cell proliferation. This assay was validated by screening a library of 1,280 bioactive molecules on both normal and tumor-derived cells where it proved more sensitive than current methods for detecting excess DNA replication. This screen identified known inducers of excess DNA replication, such as inhibitors of microtubule dynamics, and novel compounds that induced excess DNA replication in both normal and cancer cells. In addition, two compounds were identified that induced excess DNA replication selectively in cancer cells and one that induced endocycles selectively in cancer cells. Thus, this assay provides a new approach to the discovery of compounds useful for investigating the regulation of genome duplication and for the treatment of cancer.Mol Cancer Res; 9(3); 294–310. ©2011 AACR.