Fig. 4. Migration inhibitors sensitize actively migrating but not migration-restricted cells to apoptosis. (A) Cells were seeded onto slides coated with 10 µg/ml laminin and allowed to adhere. The media was exchanged for media containing migration inhibitors, cells were incubated for 24 hours and the migration rate was determined as described in Materials and Methods. (B) Cells were plated onto slides coated with 10 µg/ml laminin, allowed to migrate for 24 hours then pretreated with migration inhibitors or solvent control for 2 hours in serum-free media. Media was replaced with fresh serum-free media containing migration inhibitors or solvent control plus 1 µM camptothecin (Cpt) for SF767 cells or 100 ng/ml Trail for T98G cells. Cells were then incubated with camptothecin plus inhibitors or solvent control for 24 hours or Trail plus inhibitors or solvent control for 16 hours, fixed, stained with DAPI and apoptotic and total nuclei quantitated. Gray bars represent cells treated with camptothecin or Trail alone and black bars represent cells treated with migration inhibitor plus camptothecin or Trail. Significance with unpaired Student's t test: *P<0.05 vs camptothecin only treatment of rim cells; **P<0.001 vs Trail only treatment of rim cells. (C) Migration rate vs % apoptosis was plotted (r²=0.79) using data from Fig. 3 () and Fig. 4A,B (). Data from anti-ß1 integrin treatment of SF767 cells was deleted from this analysis.

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