Aza-Blanc Pedro, Cooper Christopher L, Wagner Klaus, Batalov Serge, Deveraux Quinn L, Cooke Michael P
Genomics Institute of the Novartis Research Foundation, 10675 John J. Hopkins Drive, San Diego, CA 92121, USA.
Mol Cell. 2003 Sep;12(3):627-37. doi: 10.1016/s1097-2765(03)00348-4.
New opportunities in mammalian functional genomics are emerging through the combination of high throughput technology and methods that allow manipulation of gene expression in living cells. Here we describe the application of an RNAi-based forward genomics approach toward understanding the biology and mechanism of TRAIL-induced apoptosis. TRAIL is a TNF superfamily member that induces selective cytotoxicity of tumor cells when bound to its cognate receptors. In addition to detecting well-characterized genes in the apoptosis pathway, we uncover several modulators including DOBI, a gene required for progression of the apoptotic signal through the intrinsic mitochondrial cell death pathway, and MIRSA, a gene that acts to limit TRAIL-induced apoptosis. Moreover, our data suggest a role for MYC and the WNT pathway in maintaining susceptibility to TRAIL. Collectively, these observations offer several insights on how TRAIL mediates the selective killing of tumor cells and demonstrate the utility of large-scale RNAi screens in mammalian cells.
高通量技术与能够在活细胞中操纵基因表达的方法相结合，为哺乳动物功能基因组学带来了新机遇。在此，我们描述了一种基于RNA干扰的正向基因组学方法在理解TRAIL诱导的细胞凋亡生物学及机制方面的应用。TRAIL是肿瘤坏死因子超家族成员，当其与同源受体结合时可诱导肿瘤细胞的选择性细胞毒性。除了检测凋亡途径中特征明确的基因外，我们还发现了几种调节因子，包括DOBI（一种通过内在线粒体细胞死亡途径促进凋亡信号传导所需的基因）和MIRSA（一种限制TRAIL诱导的细胞凋亡的基因）。此外，我们的数据表明MYC和WNT信号通路在维持对TRAIL的敏感性中发挥作用。总体而言，这些观察结果为TRAIL如何介导肿瘤细胞的选择性杀伤提供了一些见解，并证明了大规模RNA干扰筛选在哺乳动物细胞中的实用性。
