Figure 6. Versican G3 enhances tumor cell motility through EGFR signaling.

(a) The G3- and vector-transfected 66c14 cells (2×10⁵) were inoculated in 6-well culture dishes and cultured for 12 h. Monolayer G3- and vector-transfected cells were wounded by a sterile pipette tip to create a 1-mm cell-free path, washed with PBS, and then cultured in 10% FBS/DMEM medium with 2 mM hydroxyurea. These samples were also treated with or without 20 ng/ml EGF, 2.0 µM 1478PD, 50 µM PD 98059. G3-transfected cells exhibited enhanced migratory capacity, which was prevented by EGFR inhibitor AG 1478, but not by PD 98059. (b) The distances between the wounding centre and the front of the migrating cells (vertical axis) were measured for statistical analysis. (All groups compared with vector control cells, n = 10, * p<0.05, ** p<0.01, analyzed with t-test) (c) The chemotactic motility assays were performed using 24-well cell culture plates and the 3.0 µm cell culture insert. The results from a representative experiment are shown. (d) The G3-transfected 66c14 cells showed enhanced migratory capacity toward the mouse bone stromal cells that was also prevented by EGFR inhibitor AG 1478, but not by PD 98059. The modified chemotactic Boyden chamber motility assays were performed four times and were counted in three fields of views/membrane.