Discovery of novel inhibitors of ribosome biogenesis by innovative high throughput screening strategies.
Discovery of novel inhibitors of ribosome biogenesis by innovative high throughput screening strategies.
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DOI：
10.1042/bcj20190207
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发表时间：
2019-08
期刊：
影响因子：
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通讯作者：
Catherine E. Scull;Yinfeng Zhang;Nichole A. Tower;L. Rasmussen;I. Padmalayam;Robert Hunter;L. Zhai
中科院分区：
文献类型：
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作者：
Catherine E. Scull;Yinfeng Zhang;Nichole A. Tower;L. Rasmussen;I. Padmalayam;Robert Hunter;L. Zhai
在过去的二十年中，核糖体生物发生已成为癌症治疗的一个有吸引力的靶标。在这项研究中，使用两种高通量筛选来鉴定核糖体生物发生抑制剂。我们的初步筛选利用 HaloTag 选择性标记策略来识别降低 A375 恶性黑色素瘤细胞中新合成核糖体丰度的化合物。该筛选识别出 5,786 个命中化合物。使用二次筛选测试了这些初始命中化合物的子集，该二次筛选使用定量 RT-PCR 直接测量前核糖体 RNA (pre-rRNA) 丰度作为 rRNA 合成率的报告。从二次筛选中，我们鉴定出两种结构相关的化合物，它们是 rRNA 合成的有效抑制剂。这两种化合物，核糖体生物发生抑制剂 1 和 2（RBI1 和 RBI2），可导致 A375 细胞活力大幅下降，与之前发表的核糖体生物发生抑制剂 CX-5461 相当。不依赖贴壁的细胞生长测定进一步证实 RBI2 抑制细胞生长和增殖。因此，RBI 化合物作为潜在的癌症化疗药物具有进一步开发的前景。
Over the past two decades ribosome biogenesis has emerged as an attractive target for cancer treatment. In this study, two high-throughput screens were used to identify ribosome biogenesis inhibitors. Our primary screen made use of the HaloTag selective labeling strategy to identify compounds that decreased the abundance of newly synthesized ribosomes in A375 malignant melanoma cells. This screen identified 5,786 hit compounds. A subset of those initial hit compounds were tested using a secondary screen that directly measured pre-ribosomal RNA (pre-rRNA) abundance as a reporter of rRNA synthesis rate, using quantitative RT-PCR. From the secondary screen we identified two structurally related compounds that are potent inhibitors of rRNA synthesis. These two compounds, Ribosome Biogenesis Inhibitors 1 and 2 (RBI1 and RBI2), induce a substantial decrease in the viability of A375 cells, comparable to the previously published ribosome biogenesis inhibitor CX-5461. Anchorage independent cell growth assays further confirmed that RBI2 inhibits cell growth and proliferation. Thus, the RBI compounds have promising properties for further development as potential cancer chemotherapeutics.