Study on Anti-tumor Activity of Mitissimol D in vitro

To study the anti-tumor activity of mitissimol D in vitro. Hela cells were resuscitated and cultivated with routine methods. Log phase Hela cells were adjusted to 2x10(4)/mL and were seeded in 96-well plate(90 mu L/well). 100, 10, 1, 0.1, 0.01 mu g/mL mitissimol D was applied at 24h after cultured at 37 degrees C, 5% CO2. The proliferation inhibition rate and IC50 of Hela were measured by MTT assay in vitro at 24h,48h and 72h post-treated by every dose of mitissimol D respectively. DDP acted as positive control and DMSO was negative control in this experiment. The apoptosis ratio and cell-cycle of Hela cells were detected using PI stain by flow cytometry at 24h,48h and 72h treated by 100, 10, 1, 0.1 mu g/mL mitissimol D. The results indicated that mitissimol D could inhibit the proliferation of Hela cells, which the IC50 was 32.93 mu g/mL,18.49 mu g/mL,14.05 mu g/mL at 24h, 48h and 72h respectively after treated by mitissimol D. We also demonstrated that every dose of mitissimol D could induce apoptosis of Hela cells and cell apoptotic rates were 2.17%, 4.1%, 8.75% at 24h, 5.3%, 5.5%, 23.3% at 48h, 10.43%, 20.22%, at 72h after treated by mitissimol D on centrations of 1, 10, 100 mu g/mL respectively, which were significantly higher than those of the control group1.74%, 3.58% and 4.98% (P < 0.05 or P < 0.01). The analysis of cell-cycle reveals that mitissimol D can block the cell cycle into S and G2 phase. These date suggest that mitissimol D could inhibit the proliferation of Hela cells, which are associated with inducing apoptosis and blocking cell-cycle.