@article{oai:niigata-u.repo.nii.ac.jp:00006275,
 author = {CHIBA, Hitomi and KAZAMA, Takashi and YAMAMOTO, Yoko and SUYAMA, Takayuki and ITO, Masaaki},
 issue = {1},
 journal = {Acta medica et biologica, Acta medica et biologica},
 month = {Mar},
 note = {We investigated the effects of epidermal growth factor (EGF) on cell migration, and the association of focal adhesion kinase (FAK) and paxillin to cytoskeleton. A cell line of squamous cell carcinoma (SCC) of the uterine cervix, named Caski, was used. We examined the effects of different concentrations of EGF (0, 1, 10 and 100 ng/ml) on cell migration. Wound closure assay showed that cell migration within 4 h had a biphasic dose-response curve on the concentration of EGF, and that cell migration was stimulated at 1 and 10 ng/ml, with a maximum at 10 ng/ml, and suppressed at 100 ng/ml among the concentrations examined. Immunoblot and immunoprecipitation assays of whole cell lysates obtained at 20 min after the addition of EGF demonstrated that the phosphotyrosine level of FAK and paxillin had also a biphasic response curve dependent on EGF concentration, and a maximum level was observed at 10 ng/ml among the concentrations employed. To examine the amount of cytoskeleton-associated fractions of these proteins, they were extracted after a rinse in non-ionic detergent, and analyzed by immunoblot assay. At 30 min the amount of FAK and paxillin did not differ among the concentrations examined. At 90 min, however, the amount of both proteins revealed a reverse biphasic dose-response curve, with a maximal reduction at 10 ng/ml among the concentrations examined. Integrins and actinin were not changed at each concentration of EGF examined. Immunofluorescence microscopy showed that at 30 min the expression of actin stress fibers (ASF) did not differ among cells treated with each concentration of EGF. At 90 min the ratio of cells that expressed a small number of ASF and focal adhesions (FA) was significantly increased at 10 ng/ml. Although the ratio of cells that expressed large numbers of these structures was not significantly altered, ASF were gathered at the center of cells and tangled at 100 ng/ml. These data suggest that the modulation of cytoskeletal organization by EGF plays a role in the EGF-induced stimulation of cell migration.},
 pages = {31--41},
 title = {Effects of EGF on Cell Migration, and the Association of FAK and Paxillin to the Cytoskeleton},
 volume = {51},
 year = {2003}
}