National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892-2753, United States.
Methods. 2012 Jun;57(2):234-48. doi: 10.1016/j.ymeth.2012.03.031. Epub 2012 Apr 5.
High-throughput screening (HTS) provides a rapid and comprehensive approach to identifying compounds that target specific biological processes as well as genes that are essential to those processes. Here we describe a HTS assay for small molecules that induce either DNA re-replication or endoreduplication (i.e. excess DNA replication) selectively in cells derived from human cancers. Such molecules will be useful not only to investigate cell division and differentiation, but they may provide a novel approach to cancer chemotherapy. Since induction of DNA re-replication results in apoptosis, compounds that selectively induce DNA re-replication in cancer cells without doing so in normal cells could kill cancers in vivo without preventing normal cell proliferation. Furthermore, the same HTS assay can be adapted to screen siRNA molecules to identify genes whose products restrict genome duplication to once per cell division. Some of these genes might regulate the formation of terminally differentiated polyploid cells during normal human development, whereas others will prevent DNA re-replication during each cell division. Based on previous studies, we anticipate that one or more of the latter genes will prove to be essential for proliferation of cancer cells but not for normal cells, since many cancer cells are deficient in mechanisms that maintain genome stability.
高通量筛选（HTS）提供了一种快速而全面的方法，可以识别针对特定生物过程的化合物，以及对这些过程至关重要的基因。在这里，我们描述了一种用于筛选小分子的 HTS 测定法，该测定法可选择性地诱导源自人类癌症的细胞中的 DNA 再复制或内复制（即过量的 DNA 复制）。这些分子不仅将有助于研究细胞分裂和分化，而且它们可能为癌症的化学疗法提供一种新方法。由于 DNA 再复制的诱导导致细胞凋亡，因此在正常细胞中不诱导 DNA 再复制而选择性地在癌细胞中诱导 DNA 再复制的化合物可以在体内杀死癌症而不会阻止正常细胞的增殖。此外，相同的 HTS 测定法可以适应筛选 siRNA 分子，以鉴定其产物限制基因组复制到每次细胞分裂一次的基因。这些基因中的一些可能在正常的人类发育过程中调节终末分化的多倍体细胞的形成，而其他基因将在每次细胞分裂过程中阻止 DNA 再复制。根据先前的研究，我们预计其中一个或多个后者的基因将被证明对于癌细胞的增殖是必需的，但对于正常细胞则不是必需的，因为许多癌细胞缺乏维持基因组稳定性的机制。
