Effects Of Casticin On Proliferation And Apoptosis Of Human Ovarian Cancer HO-8910 Cells In Vitro

Objective:To investigate the effects of CAS (5, 3′-dihydroxy-3, 6, 7, 4′-tetram thoxyflavone; casticin) on proliferation and apoptosis of human ovarian cancer HO-8910 cells in vitro, and its mechanisms.Methods:The inhibitory effect of CAS on the proliferation of human ovarian cancer HO-8910 cells was evaluated by the MTT assay and the apoptotic effect was demonstrated by Hoechst33258 staining and flow cytometric analysis. The expression of proteins was analyzed by Western blot.Results:MTT assay showed that that CAS inhubited the growth of HO-8910 cells, in a dose- and time-dependent manner. CAS(2, 4, 8μg/ml) for 48 h suppressed HO-8910 cells growth by 8.4%±1.8%, 28.6%±2.1%, 35.1%±3.1%, and the IC50 was 6.46μg/mL.The typical morphological change of apoptosis, such as nuclear fragmentation, nuclear karyopycnosis, edge set of nuclear chromosomes, the formation of apoptotic bodies, were observed using fluorescence microscope after exposure of HO-8910 cells to 2, 4, 8μg/mL CAS for 48 h.Flow cytometry analysis revealed that treatment HO-8910 cells with CAS(2, 4, 8μg/ml) for 48 h, the percentage of apoptotic cells were 20.3%,33.4%,52.8% respectively, these results showed that CAS could induce apoptosis of HO-8910 cells in a dose-dependece manner.There were singnificantly difference between HO-8910 cells treated with 2, 4, 8μg/mL CAS and the normal HO-8910 cells cultrued with medium for 48 h in the mean relative density of expression of cyclin B1 protein (P<0.05).The relative density were 1.000±0.010, 0.580±0.060, 0.590±0.03, 0.200±0.045 respectively.The relative density of p21 protein were 1.000±0.120, 3.950±0.150, 4.790±0.420, 8.220±0.580 respectively; and the relative density of caspse-3 protein were 1.000±0.065, 1.450±0.071, 2.770±0.270, 2.650±0.150 respectively. Western blot showed that the inhibition of proliferation and induction of apoptosis of CAS may be through upregulation of expression of caspse-3 protein and p21 protein and depression of cyclin B1 protein in human ovarian cancer HO-8910 cells.Conclusions:CAS could inhibit proliferation and induce apoptosis of human ovarian cancer HO-8910 cells in vitro through downregulation of cyclin B1 expression and activation of p21 and caspase-3.