Figure 4.

Overexpression of FoxM1 promoted growth and clonogenicity and induced invasion and migration in CRC cells. A: FoxM1 cDNA was transfected in HCT-15 and LOVO and stable clones were selected. HCT-15 and LOVO cells with vector alone or FoxM1-transfected clones were lysed and proteins were immunoblotted with FoxM1 and β-actin. B: Vector and FoxM1-overexpressing HCT-15 clones 1 and 3 and LOVO clones 1 and 2 were seeded in 96-well plates. After 48 hours, cell proliferation assays were performed using MTT as described in Materials and Methods. Data are reported as means ± SD of three independent experiments, with replicates of six wells for all of the doses and vehicle control for each experiment *P < 0.05, Student's t-test. C: Clonogenic assays were performed using vector and FoxM1-overexpressing HCT-15 clones 1 and 3 as described in Materials and Methods (upper panel). HCT-15 vector and HCT-15 clones 1 and 3 were plated in soft agar plates for 4 weeks, after which cells were stained and manually counted (lower panel). *P < 0.05, Student's t-test. D: HCT-15 and LOVO cells with vector alone or FoxM1-transfected clones were lysed and proteins were immunoblotted with MMP-2, MMP-9, and β-actin. E: Invasion-migration assays were performed using vector and FoxM1-overexpressing HCT-15 clones 1 and 3 as described in Materials and Methods. F: Manual counts of invasion and migration experiment of HCT-15 vector, clone 1, and clone 3 cells treated as just described for (E). *P < 0.005.