title: Identification of kinases and regulatory proteins required for cell migration using a transfected cell-microarray system.
abstract: BACKGROUND
  Cell migration plays a major role in a variety of normal biological processes, and a detailed understanding of the associated mechanisms should lead to advances in the medical sciences in areas such as cancer therapy. Previously, we developed a simple chip, based on transfected-cell microarray (TCM) technology, for the identification of genes related to cell migration. In the present study, we used the TCM chip for high-throughput screening (HTS) of a kinome siRNA library to identify genes involved in the motility of highly invasive NBT-L2b cells.
  
  
  RESULTS
  We performed HTS using TCM coupled with a programmed image tracer to capture time-lapse fluorescence images of siRNA-transfected cells and calculated speeds of cell movement. This first screening allowed us to identify 52 genes. After quantitative PCR (qPCR) and a second screening by a conventional transfection method, we confirmed that 32 of these genes were associated with the migration of NBT-L2b cells. We investigated the subcellular localization of proteins and levels of expression of these 32 genes, and then we used our results and databases of protein-protein interactions (PPIs) to construct a hypothetic but comprehensive signal network for cell migration.
  
  
  CONCLUSIONS
  The genes that we identified belonged to several functional categories, and our pathway analysis suggested that some of the encoded proteins functioned as the hubs of networks required for cell migration. Our signal pathways suggest that epidermal growth factor receptor (EGFR) is an upstream regulator in the network, while Src and GRB2 seem to represent nodes for control of respective the downstream proteins that are required to coordinate the many cellular events that are involved in migration. Our microarray appears to be a useful tool for the analysis of protein networks and signal pathways related to cancer metastasis.
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. r-nagasaki@aist.go.jp.|Research Fellow of Japan Society for the Promotion of Science, 5-3-1 Kojimachi, Chiyoda-ku, Tokyo, 102-0083, Japan. r-nagasaki@aist.go.jp.
authors: Reiko
authors: R
authors: Onuki-Nagasaki
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. a-nagasaki@aist.go.jp.
authors: Akira
authors: A
authors: Nagasaki
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. Hakamada.Kazumi@sysmex.co.jp.|Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka, 560-8531, Japan. Hakamada.Kazumi@sysmex.co.jp.|Current address: Central Research Laboratories Sysmex Corporation, 4-4-4 Takatsukadai, Nishi-ku, Kobe, 657-2271, Japan. Hakamada.Kazumi@sysmex.co.jp.
authors: Kazumi
authors: K
authors: Hakamada
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. t-uyeda@aist.go.jp.
authors: Taro Q P
authors: TQ
authors: Uyeda
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. masato-miyake@aist.go.jp.
authors: Masato
authors: M
authors: Miyake
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. jun_miyake@bpe.es.osaka-u.ac.jp.|Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka, 560-8531, Japan. jun_miyake@bpe.es.osaka-u.ac.jp.
authors: Jun
authors: J
authors: Miyake
authors: Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8566, Japan. s-fujita@aist.go.jp.
authors: Satoshi
authors: S
authors: Fujita
mesh_terms: D002465:Cell Movement; D015723:Gene Library; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D009362:Neoplasm Metastasis; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology*; D010770:Phosphotransferases / Q000032:analysis*; D034741:RNA, Small Interfering; D015398:Signal Transduction; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering; D010770:Phosphotransferases
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