Figure 3
SPC-A-1sci cells acquire increased potentials of migration and invasion. A, In vitro growth curves of SPC-A-1sci and SPC-A-1 cells, cells (2 × 104/well) were cultured in 24-well plates, after which cells were trypsined and counted at indicated times. Results are expressed as mean ± SD; *, P < 0.05. B, Soft agar colony formation of SPC-A-1sci and SPC-A-1 cells, cells (1 × 103/well) were cultured in 24-well plates, colonies were photographed and counted after three weeks. Results are expressed as mean ± SD; *, P < 0.05. C, In vivo growth curve of SPC-A-1sci and SPC-A-1 cells, cells (2.0 × 106 per mouse) were s.c. implanted into NOD/SCID mice, after which the tumour volume was measured over time. Results are expressed as mean ± SD. C, Motility activities of SPC-A1sci and SPC-A-1 cells were determined by wound healing assays Confluent monolayers of SPC-A1sci and SPC-A-1 cells in fibronectin coated 24-well plates were wounded by scratching, after which the wells were washed, incubated with conditioned medium for 24 h and photographed (lower). Motility was assessed on the basis of percentage of wounded area filled in (upper). Results are expressed as mean ± SD; **, P < 0.01. D and E, Trans-well migration and invasion assays of SPC-A-1sci and SPC-A-1 cells For migration assay (D), cells (2.5 × 104/well) were seeded into non-coated trans-well plates and culture for 16 h at 37°C; for invasion assays (E), cells (1.0 × 105/well) were seeded into Matrigel-coated trans-well plates and cultured for 24 h at 37°C, after which cells that had migrated or invaded to the underside of the inserts were stained with H&E and the cells on each insert were photographed (lower) and quantified (upper) at 400× magnification. All experiments were repeated thrice; Results are expressed as mean ± SD; *, P < 0.05.