Inhibitory Effects On Tca8113 Cell Proliferation And Migration Of PDE And VP-16 In Different Concentration Ranges

Objective To investigate the inhibitory effects on Tca8113 cell proliferation and migration of 4-p-amino-benzoinc acid-4’-demethylepipodophyllotoxin ester(PDE)and VP-16 in Different Concentration Ranges.Methods Sulforhodamine B assay was used to examine the inhibitory effect of PDE and VP-16 against Tca8113 cells.The influence of cell cycle in Tca8113 cells was analyzed by Flow Cytometer.Tca8113 cell migration was measured by transwell chamber migration assay.The activities of matrix metalloproteinase MMP-2 and MMP-9 were assessed by Gelatin Zymography.Results SRB assay showed that PDE and VP-16 inhibited proliferation of Tca8113 cells(P<0.05)in both of high concentration group(12.5-200 μg/mL)and low concentration group(0.05-0.8 μg/mL).VP-16 inhibited proliferation of Tca8113 cells stronger than that of PDE in high concentration group(12.5～200 μg/mL).However,PDE inhibited proliferation of Tca8113 cells stronger than that of VP-16 in low concentration group(0.05～0.8 μg/mL).After treatment with high concentration group(25～200μg/mL)of PDE at 24h,Flow cytometry results showed that the cell cycle was arrested at the G2/M phase with the concentration between 25～100 μg/mL,but the cell cycle was arrested at the S phase with the concentration 200 μg/mL treatment.After treatment with low concentration group(0.1～0.8μg/mL)of PDE at 24h,the cell cycle was arrested at the G2/M phase with 0.8 μg/mL treatment.The cell cycle was arrested at G0/G1 phase after treatment with high concentration group(25～200μg/mL)of VP-16 at 24h,while the cell cycle was arrested at the G2/M phase and S phase after treatment with low concentration group(0.1～0.8μg/mL)of VP-16 at 24h.Transwell chamber assay showed that PDE inhibited Tca8113 cell migration in both high concentrations group(3.125～25μg/mL)and low concentrations group(0.025～0.2μg/mL).The migration inhibition rates of PDE(3.125～25μg/mL)treatment were 10.8%、28.5%、38.9%and 57.3%.The migration inhibition rates treated with PDE(0.025～0.2 μg/mL)were 6.11%.11.6%、20.1%and 33.6%.As same as PDE,VP-16 also inhibited Tca8113 cell migration in both high concentrations group(3.125～25μgg/mL)and low concentrations group(0.025～0.2μg/mL).After treatment with VP-16(3.125～25μg/mL),the migration inhibition rates were 27.4%,42.8%,49.3%and 42.8%.Treat with VP-16(0.025～0.2 μg/mL),the migration inhibition rates were 1.71%,25.3%,21.1%and 23.3%.Gelatin Zymography results showed that PDE markedly inhibited the activities of MMP-2 and MMP-9 in culture supernatant of Tca8113 cells in both high concentrations group(25～200μg/mL)and low concentrations group(0.1～0.8μg/mL).With the high concentrations of PDE(25～200μg/mL),the inhibition rates to MMP-2 were 12.5%、19.5%、25.7%、38.0%,and the inhibition rates to MMP-9 were 28.6%、32.7%、44.0%、46.7%.With the low concentrations of PDE(0.1～0.8μg/mL),the inhibition rates to MMP-2 were 23.5%、34.1%、28.8%、44.8%,and the inhibition rates to MMP-9 were 11.0%、31.2%、29.1%、36.8%.However,VP-16 inhibited the activities of MMP-2 and MMP-9 weakly in both high concentrations group(25～200pμg/mL)and low concentrations group(0.1～0.8 μg/mL).With the high concentrations of VP-16(25～200μg/mL),the inhibition effects to MMP-2 and MMP-9 were not significant.With the low concentrations of VP-16(0.1～0.8μg/mL),the inhibition rates to MMP-2 were 11.7%,15.3%,15.8%and 12.6%,and the inhibition rates to MMP-9 were 9.49%,10.3%,13.6%and 18.6%.Conclusion PDE and VP-16 inhibited proliferation and migration of Tca8113 cells(P<0.05)in both of high concentration group and low concentration group.There are different effects on cell cycle after treatment with PDE and VP-16 in both high concentration group and low concentration group.VP-16 inhibited the activities of MMP-2 and MMP-9 weakly in both high concentrations group and low concentrations group.