The Combination Of Sorafenib And Sodium Cantharidinate Procides Anti-Proliferative Effects In Liver Cancer HepG2 Cell

ObjectiveTo investingate the anti-proliferative effect of the combination of Sorafenib and sodium cantharidinate for liver cancer HepG2 cell and the relative machanisms.MethodsThe study included the group of sorafenib, sodium cantharidinate alone and their combination and control, the 50% inhibiting concentration (IC50) of Sorafenib and sodium cantharidinate were acquired by the CCK-8 assays? and the concentration of Sorafenib and sodium cantharidinate for aftermentioned study was designated depending on the IC50, The antitumor effect of sorafenib and sodium cantharidinate for HepG2 cell in vitro was detected by CCK-8 assays, the effect of different inhibitor concentrations was assessed by plate clone formation asssy;the flow cytometry was used to examine the cell cycle. Caspase 3 Activity Colorimetric Assay was used to test the activity level of Caspace 3.the western bloting was used to examine the expressions of the ERK and pERKResultsBoth sorafenib and sodium cantharidinate can inhibit the proliferation of HepG2, the dose-time dependent effect was showed in the study, and revealing a synergistic effect of antitumor when the two agents were combined used(p<0.05), and revealed a synergistic effect of antitumor when the two agents were combined used in concentration of 4+3.88μmol/L at 48h time point, most of others showed the additive effect; The plate colony-forming asssy showed rate of clone formation of sorafenib or sodium cantharidinate alone or their combination were lower with higher of dose, There showed the dose-dependent decrease in colony-forming ability, the group of combined treatment had the significantly lower rate of clone formation than others(p<0.05);The rate of cell arrested the cell cycle at G1 phases of sorafenib and sodium cantharidinate alone or their combination were64.86±1.44%、 60.68±1.48%、69.13±1.85%, which were higher than the control group’s49.89±3.30%(F=45.936,p<0.001), and the group of combined treatment had significantly higher rate than the groups of alone(p<0.05); The activity level of Caspase-3 were increased in the group of sorafenib or sodium cantharidinate alone or their combination,and the level of combination was highest(p<0.05).The alone or combined use of the two agents didn’t show a significant effect for the expression of ERK compared with the control (F=2.928, P=0.100),But for p-ERK,the related expressions of the group of sorafenib alone and combined treatment were 0.3122±0.0180及0.3714± 0.0444,which were lower than the control’s 0.4694±0.0568(p<0.05), whereas the group of sodium cantharidinate alone was 0.5718±0.0503,which was higher than the control(p=0.023)Conclusionthe combination of sorafenib and sodium cantharidinate show a synergist anti-proliferative effect for HepG2 cell,the mechanisms of the effect may be related with the arrest of cell cycle,activate the caspase 3 and the inhibition of the RAF/MEK/ERK signaling pathway.