Figure 2

Expression of NPM1 was elevated in highly invasive colon cancer cell line HCT116. A. Western blotting analysis of NPM1 expression in five different colon cancer cell lines with distinct invasive potentials; β-actin was used as a loading control. B. Cell proliferation was measured by the BrdU incorporation assay. The proliferation rates of HCT116 and Colo205 were significantly higher than those of HT29. *** P < 0.0001, ** P < 0.001. C. Immunocytochemical staining of PCNA in different cell lines. Expression of PCNA was higher in the HCT116 and Colo205 cell lines than in the HT29 cell line. Picture was taken with a light microscope at 400 magnifications. *** P < 0.0001. D. HCT116 and Colo205 cells exhibited higher invasive ability than HT29 cells. 5 × 10⁵ cells in 400 ul serum free medium were loaded into transwell inserts with 8μm pore polycarbonate filters which coated with matrigel. After 48h incubation, the invaded cells on the bottom side of the membrane were fixed and stained with 0.005% crystal violet. Then,the number of invaded cells was counted in three randomly chosen areas (400 × magnification) and summarized in the right panel. *** P < 0.0001, ** P < 0.001. E. HCT116 and Colo205 cells exhibited higher migration ability than HT29 cells. Migration assay was performed the same as transwell invasion assay except that there was no coating matrigel on the filters. *** P < 0.0001. F. Representative images of wound healing assays at 0 and 24 h (100 × migration). The migration distance was quantified by subtracting the width of the wounds at each time point from the width of the initial wounds. HCT116 cells showed increased migration ability compared with the other cell lines. *** P < 0.0001.