Whitehurst Angelique W, Bodemann Brian O, Cardenas Jessica, Ferguson Deborah, Girard Luc, Peyton Michael, Minna John D, Michnoff Carolyn, Hao Weihua, Roth Michael G, Xie Xian-Jin, White Michael A
Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
Nature. 2007 Apr 12;446(7137):815-9. doi: 10.1038/nature05697.
Abundant evidence suggests that a unifying principle governing the molecular pathology of cancer is the co-dependent aberrant regulation of core machinery driving proliferation and suppressing apoptosis. Anomalous proteins engaged in support of this tumorigenic regulatory environment most probably represent optimal intervention targets in a heterogeneous population of cancer cells. The advent of RNA-mediated interference (RNAi)-based functional genomics provides the opportunity to derive unbiased comprehensive collections of validated gene targets supporting critical biological systems outside the framework of preconceived notions of mechanistic relationships. We have combined a high-throughput cell-based one-well/one-gene screening platform with a genome-wide synthetic library of chemically synthesized small interfering RNAs for systematic interrogation of the molecular underpinnings of cancer cell chemoresponsiveness. NCI-H1155, a human non-small-cell lung cancer line, was employed in a paclitaxel-dependent synthetic lethal screen designed to identify gene targets that specifically reduce cell viability in the presence of otherwise sublethal concentrations of paclitaxel. Using a stringent objective statistical algorithm to reduce false discovery rates below 5%, we isolated a panel of 87 genes that represent major focal points of the autonomous response of cancer cells to the abrogation of microtubule dynamics. Here we show that several of these targets sensitize lung cancer cells to paclitaxel concentrations 1,000-fold lower than otherwise required for a significant response, and we identify mechanistic relationships between cancer-associated aberrant gene expression programmes and the basic cellular machinery required for robust mitotic progression.
大量证据表明，癌症分子病理学的一个统一原则是驱动增殖和抑制凋亡的核心机制的相互依赖的异常调节。参与支持这种致瘤调节环境的异常蛋白很可能是异质性癌细胞群体中的最佳干预靶点。基于RNA介导的干扰（RNAi）的功能基因组学的出现，为在预先设定的机制关系概念框架之外，获得支持关键生物系统的经过验证的基因靶点的无偏倚综合集合提供了机会。我们将基于细胞的高通量单孔/单基因筛选平台与化学合成的小干扰RNA的全基因组合成文库相结合，以系统地探究癌细胞化疗反应性的分子基础。在一项依赖紫杉醇的合成致死筛选中，使用了人非小细胞肺癌细胞系NCI-H1155，旨在识别在亚致死浓度的紫杉醇存在下能特异性降低细胞活力的基因靶点。使用严格的客观统计算法将错误发现率降低到5%以下，我们分离出一组87个基因，这些基因代表了癌细胞对微管动力学废除的自主反应的主要焦点。在这里，我们表明，这些靶点中的几个能使肺癌细胞对紫杉醇的敏感性提高到比显著反应所需浓度低1000倍的水平，并且我们确定了癌症相关异常基因表达程序与稳健有丝分裂进展所需的基本细胞机制之间的机制关系。
