A genome-wide RNAi screen identifies multiple RSK-dependent regulators of cell migration
A genome-wide RNAi screen identifies multiple RSK-dependent regulators of cell migration
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DOI：
10.1101/gad.1989110
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发表时间：
2010-12-01
影响因子：
10.5
通讯作者：
Haber, Daniel A.
中科院分区：
文献类型：
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作者：
Smolen, Gromoslaw A.;Zhang, Jianmin;Haber, Daniel A.
为了确定调节上皮细胞迁移的功能途径，我们使用55,000个汇集的慢病毒shrna进行了全基因组RNAi筛选，靶向类似11,000个基因，选择具有增加运动性的转导细胞。严格的验证方案产生了一组31个基因，代表不同的途径，敲除显著增强细胞迁移。其中一些通路具有上皮-间质转化（EMT）的特征，它们共同涉及转录、细胞信号传导和代谢的关键调节因子，以及细胞运输的新调节剂，如DLG5。在描述介导这些迁移表型的下游途径时，我们观察到erk的普遍激活及其对RSK效应物的深刻依赖。药理抑制RSK可显著抑制所有31个基因敲低诱导的上皮细胞迁移，这表明多种迁移途径在该激酶上的趋同可能为包括癌症转移在内的细胞迁移障碍提供治疗机会。
To define the functional pathways regulating epithelial cell migration, we performed a genome-wide RNAi screen using 55,000 pooled lentiviral shRNAs targeting similar to 11,000 genes, selecting for transduced cells with increased motility. A stringent validation protocol generated a set of 31 genes representing diverse pathways whose knockdown dramatically enhances cellular migration. Some of these pathways share features of epithelial-to-mesenchymal transition (EMT), and together they implicate key regulators of transcription, cellular signaling, and metabolism, as well as novel modulators of cellular trafficking, such as DLG5. In delineating downstream pathways mediating these migration phenotypes, we observed universal activation of ERKs and a profound dependence on their RSK effectors. Pharmacological inhibition of RSK dramatically suppresses epithelial cell migration induced by knockdown of all 31 genes, suggesting that convergence of diverse migratory pathways on this kinase may provide a therapeutic opportunity in disorders of cell migration, including cancer metastasis.