Department of Pharmacology, Brain Science and Engineering Institute, Kyungpook National University School of Medicine, Joong-gu, Daegu, Korea.
Cell Adh Migr. 2010 Apr-Jun;4(2):207-10. doi: 10.4161/cam.4.2.11073. Epub 2010 Apr 22.
Cell migration is a central process that is essential for embryonic development, wound repair, inflammatory response, homeostasis and tumor metastasis. A method of genome-wide selection based on the gain-of-function has been devised to identify novel cell migration-promoting genes in cultured cells. After the introduction of the retroviral mouse brain cDNA library into NIH3T3 mouse fibroblast cells, migration-promoted cells were selected by a three-dimensional migration assay using cell culture inserts. After five rounds of enrichment, cDNAs were retrieved from the cells that passed the selection processes. Cell migration-promoting activity was confirmed by independent migration assays for the retrieved cDNAs. Multiple cell migration-promoting genes were successfully isolated by this method. The genes identified can be used to gain a systematic view of cell migration. The gain-of-function selection method described here can be combined with RNAi-mediated loss-of-function screen or selection to be a more powerful tool for the systems biology research of cell migration.
细胞迁移是一个核心过程,对于胚胎发育、伤口修复、炎症反应、内稳态和肿瘤转移都是必不可少的。已经设计了一种基于功能获得的全基因组选择方法,以鉴定培养细胞中促进细胞迁移的新基因。将逆转录病毒小鼠脑 cDNA 文库导入 NIH3T3 小鼠成纤维细胞后,通过使用细胞培养插入物的三维迁移测定法选择促进迁移的细胞。经过五轮富集后,从通过选择过程的细胞中回收 cDNA。通过独立的迁移测定法验证回收 cDNA 的细胞迁移促进活性。通过这种方法成功分离了多个促进细胞迁移的基因。所鉴定的基因可用于对细胞迁移进行系统观察。这里描述的功能获得选择方法可以与 RNAi 介导的功能丧失筛选或选择相结合,成为细胞迁移的系统生物学研究的更强大工具。