A small interfering RNA screen for modulators of tumor cell motility identifies MAP4K4 as a promigratory kinase

被引:162
作者
Collins, CS
Hong, JY
Sapinoso, L
Zhou, YY
Liu, Z
Micklash, K
Schultz, PG
Hampton, GM
机构
[1] Novartis Res Fdn, Genom Inst, San Diego, CA 92121 USA
[2] Scripps Res Inst, Skaggs Inst Chem Biol, Dept Chem, La Jolla, CA 92037 USA
关键词
automated; RNA interference;
D O I
10.1073/pnas.0600040103
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Cell motility is a complex biological process, involved in development, inflammation, homeostasis, and pathological processes such as the invasion and metastatic spread of cancer. Here, we describe a genomic screen designed to identify inhibitors of cell migration. A library of 10,996 small interfering RNAs (targeting 5,234 human genes) was screened for their ability to block the migration of a highly motile ovarian carcinoma cell line, SKOV-3, by using a 384-well wound-healing assay coupled with automated microscopy and wound quantification. Two or more small interfering RNAs against four genes, CDK7, DYRK1B, MAP4K4 (NIK/HGK) (MAP4K4, mitogen-activated protein 4 kinase 4), and SCCA-1 (SerpinB3), potently blocked the migration of SKOV-3 cells, concordant with reduced transcript levels. Further studies of the promigratory role of MAP4K4 showed that the knockdown of this transcript inhibited the migration of multiple carcinoma cell lines, indicating a broad role in cell motility and potently suppressed the invasion of SKOV-3 cells in vitro. The effect of MAP4K4 on cellular migration was found to be mediated through c-Jun N-terminal kinase, independent of AP1 activation and downstream transcription. Accordingly, small molecule inhibition of c-Jun N-terminal kinase suppressed SKOV-3 cell migration, underscoring the potential therapeutic utility of mitogen-activated protein kinase pathway inhibition in cancer progression.
引用
收藏
页码:3775 / 3780
页数:6
相关论文
共 16 条
[1]   Identification of modulators of TRAIL-induced apoptosis via RNAi-based phenotypic screening [J].
Aza-Blanc, P ;
Cooper, CL ;
Wagner, K ;
Batalov, S ;
Deveraux, QL ;
Cooke, MP .
MOLECULAR CELL, 2003, 12 (03) :627-637
[2]   Inhibition of constitutively active Stat3 suppresses growth of human ovarian and breast cancer cells [J].
Burke, WM ;
Jin, XH ;
Lin, HJ ;
Huang, M ;
Liu, R ;
Reynolds, RK ;
Lin, JY .
ONCOGENE, 2001, 20 (55) :7925-7934
[3]   Identification of a novel inhibitor of mitogen-activated protein kinase kinase [J].
Favata, MF ;
Horiuchi, KY ;
Manos, EJ ;
Daulerio, AJ ;
Stradley, DA ;
Feeser, WS ;
Van Dyk, DE ;
Pitts, WJ ;
Earl, RA ;
Hobbs, F ;
Copeland, RA ;
Magolda, RL ;
Scherle, PA ;
Trzaskos, JM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (29) :18623-18632
[4]   SKI-606, a Src/AbI inhibitor with in vivo activity in colon tumor xenograft models [J].
Golas, JM ;
Lucas, J ;
Etienne, C ;
Golas, J ;
Discafani, C ;
Sridharan, L ;
Boghaert, E ;
Arndt, K ;
Ye, F ;
Boschelli, DH ;
Li, FB ;
Titsch, C ;
Huselton, C ;
Chaudhary, I ;
Boschelli, F .
CANCER RESEARCH, 2005, 65 (12) :5358-5364
[5]   Optimization of 2-phenylaminoimidazo[4,5-h]isoquinolin-9-ones:: Orally active inhibitors of lck kinase [J].
Goldberg, DR ;
Butz, T ;
Cardozo, MG ;
Eckner, RJ ;
Hammach, A ;
Huang, J ;
Jakes, S ;
Kapadia, S ;
Kashem, M ;
Lukas, S ;
Morwick, TM ;
Panzenbeck, M ;
Patel, U ;
Pav, S ;
Peet, GW ;
Peterson, JD ;
Prokopowicz, AS ;
Snow, RJ ;
Sellati, R ;
Takahashi, H ;
Tan, J ;
Tschantz, MA ;
Wang, XJ ;
Wang, Y ;
Wolak, J ;
Xiong, P ;
Moss, N .
JOURNAL OF MEDICINAL CHEMISTRY, 2003, 46 (08) :1337-1349
[6]   MAP kinases and cell migration [J].
Huang, C ;
Jacobson, K ;
Schaller, MD .
JOURNAL OF CELL SCIENCE, 2004, 117 (20) :4619-4628
[7]   JNK phosphorylates paxillin and regulates cell migration [J].
Huang, C ;
Rajfur, Z ;
Borchers, C ;
Schaller, MD ;
Jacobson, K .
NATURE, 2003, 424 (6945) :219-223
[8]   Disruption of basal JNK activity differentially affects key fibroblast functions important for wound healing [J].
Javelaud, D ;
Laboureau, J ;
Gabison, E ;
Verrecchia, F ;
Mauviel, A .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (27) :24624-24628
[9]   Cell migration: A physically integrated molecular process [J].
Lauffenburger, DA ;
Horwitz, AF .
CELL, 1996, 84 (03) :359-369
[10]   A conserved interaction between β1 integrin/PAT-3 and Nck-interacting kinase/MIG-15 that mediates commissural axon navigation in C-elegans [J].
Poinat, P ;
De Arcangelis, A ;
Sookhareea, S ;
Zhu, XP ;
Hedgecock, EM ;
Labouesse, M ;
Labouesse, EG .
CURRENT BIOLOGY, 2002, 12 (08) :622-631