Arlt Dorit, Huber Wolfgang, Liebel Urban, Schmidt Christian, Majety Meher, Sauermann Mamatha, Rosenfelder Heiko, Bechtel Stephanie, Mehrle Alexander, Bannasch Detlev, Schupp Ingo, Seiler Markus, Simpson Jeremy C, Hahne Florian, Moosmayer Petra, Ruschhaupt Markus, Guilleaume Birgit, Wellenreuther Ruth, Pepperkok Rainer, Sültmann Holger, Poustka Annemarie, Wiemann Stefan
Division of Molecular Genome Analysis, German Cancer Research Center, Heidelberg, Germany.
Cancer Res. 2005 Sep 1;65(17):7733-42. doi: 10.1158/0008-5472.CAN-05-0642.
Cancer transcription microarray studies commonly deliver long lists of "candidate" genes that are putatively associated with the respective disease. For many of these genes, no functional information, even less their relevance in pathologic conditions, is established as they were identified in large-scale genomics approaches. Strategies and tools are thus needed to distinguish genes and proteins with mere tumor association from those causally related to cancer. Here, we describe a functional profiling approach, where we analyzed 103 previously uncharacterized genes in cancer relevant assays that probed their effects on DNA replication (cell proliferation). The genes had previously been identified as differentially expressed in genome-wide microarray studies of tumors. Using an automated high-throughput assay with single-cell resolution, we discovered seven activators and nine repressors of DNA replication. These were further characterized for effects on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling (G1-S transition) and anchorage-independent growth (tumorigenicity). One activator and one inhibitor protein of ERK1/2 activation and three repressors of anchorage-independent growth were identified. Data from tumor and functional profiling make these proteins novel prime candidates for further in-depth study of their roles in cancer development and progression. We have established a novel functional profiling strategy that links genomics to cell biology and showed its potential for discerning cancer relevant modulators of the cell cycle in the candidate lists from microarray studies.
癌症转录微阵列研究通常会给出一长串与相应疾病可能相关的“候选”基因。对于其中许多基因，由于它们是在大规模基因组学方法中被鉴定出来的，所以尚未建立起功能信息，更不用说它们在病理状况下的相关性了。因此，需要策略和工具来区分仅仅与肿瘤相关的基因和蛋白质与那些与癌症有因果关系的基因和蛋白质。在这里，我们描述了一种功能分析方法，即我们在与癌症相关的实验中分析了103个以前未被表征的基因，这些实验探究了它们对DNA复制（细胞增殖）的影响。这些基因先前已在肿瘤的全基因组微阵列研究中被鉴定为差异表达基因。通过使用具有单细胞分辨率的自动化高通量实验，我们发现了7个DNA复制激活因子和9个DNA复制抑制因子。进一步对它们在细胞外信号调节激酶1/2（ERK1/2）信号传导（G1-S期转换）和非锚定依赖性生长（致瘤性）方面的作用进行了表征。鉴定出了一个ERK1/2激活的激活蛋白和一个抑制蛋白以及三个非锚定依赖性生长的抑制因子。肿瘤和功能分析的数据使这些蛋白质成为进一步深入研究它们在癌症发生和发展中作用的新的主要候选对象。我们建立了一种将基因组学与细胞生物学联系起来的新型功能分析策略，并展示了其在从微阵列研究的候选列表中识别细胞周期中与癌症相关的调节因子的潜力。
