| Objective: The present study aimed to study the effects of chrysin combined with diosmetin or triptolide on the proliferation,migration,apoptosis and cycle arrest of human liver cancer Hep G2 cells.To observe the inhibitory action of chrysin combined with diosmetin or triptolide on hepatoma H22 solid tumor and and evaluate their efficacy in vivo.Methods: The inhibitory effect of baicalin,baicalein,diosmetin and chrysin on human ovarian cancer cells A2780 and SKOV3,gastric cancer cells BGC-823,liver cancer cells Hep G2 were determined by MTT assay.MTT was used to evaluate the effects of chrysin combined with diosmetin or triptolide on growth,respectively,in Hep G2 cells after 24,48 and 72 h.Morphological changes accompanying Hep G2 cells growth inhibition were observed using Hoechst 33258 staining in fluorescence microscopy.Cell scratch healing experiments were used to observe the effects of chrysin combined with diosmetin or triptolide on scratch healing after 0,24 and 48 h.Annexin V-FITC/PI and PI were used as fluoresent probes to measure the apoptosis rate and cell cycle respectively with flow cytometer.H22 ascitic cells were inoculated right armpit of mice to construct solid tumor model,the mice that successful tumor types were randomly divided into eight groups.The effects of different groups on the weight of hepatoma H22 solid tumor,inhibition rate,the organ index and lymphocyte subsets were observed.Results:(1)Both chrysin and diosmetin were the most sensitive to Hep G2 cells,with IC50 values of 18.21 μmol/L and 20.94 μmol/L respectively.The results of MTT assay revealed that chrysin combined with diosmetin or triptolide markedly inhibited the growth of Hep G2 cells in a time-and dose-dependent manner,and shows a synergistic effect.(2)Triptolide and diosmetin could significantly inhibit cell migration,but the inhibitory effect of chrysin was not obvious.(3)In both the single drug group and the combination group,the phenomenon of nuclear edge consolidation,fragmentation,chromatin aggregation and apoptotic bodies were observed in Hep G2 cells,among which the combination group had the largest number of apoptotic cells.(4)Triptolide had no obvious effect on Hep G2 cells in each cycle,chrysin can induce G1 phase arrest,diosmetin can induce G2 phase arrest,triptolide/chrysin combined group can induce G1 phase and G2 phase arrest,chrysin/diosmetin combined group can induce S phase and G2 phase arrest.(5)The apoptosis rates of chrysin group,diosmetin group,triptolide group,chrysin/diosmetin combined group,triptolide/chrysin combined group were 16.39%,35.17%,5.07%,41.5%,and 24.9%,respectively.All of them were significantly higher than the normol group(3.21%),and the apoptosis rate of combination group was higher than the single drug group.(6)H22 solid tumor experment: Chrysin/diosmetin combined group had certain inhibitory effect on H22 solid tumor with an inhibition rate of 38.83%,which had a certain protective effect on the immune system.Triptolide/chrysin combined group had no significant inhibitory effect on tumor growth and no significant regulatory effect on immune system.Conclusion: The combined applications of chrysin/diosmetin and triptolide/chrysin had a stronger effect on the proliferation,migration,cell cycle and apoptosis of Hep G2 cells than that of single drug.H22 solid tumor experment showed that the combined application of chrysin/diosmetin had a stronger inhibitory effect on H22 solid tumors than single drug application. |
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