Effect Of Norcantraridin Combined With Evodiamine On HepG2Cell Lines In Vitro

Objective To investigate the combined effect of evodiamine andnorcantharidin on liver cancer HepG2cells in vitro.Method The inhibitory effect of evodiamine or/and norcantharidin wasdetermined by MTT assay. The combined effect of evodiamine or/andnorcantharidin on cell cycle, cell apoptosis and activity of axygen wereassessed by flow cytometry (FCM). The expressions of Bax and Bcl-2wereexamined by Western blotting.Results1. The growth of Hepg2cell was inhibited by NCTD or EVO indifferent concentration. The inhibitory effect of drug was in a dose and timedependent way. MTT results showed that the IC50of NCTD were177.6,102.1,76.3μmol/L and EVO were129.9,20.9,9.3μmol/L in24h,48h,72h respectively.2. EVO could enhance the inhibitory effect of NCTD on HepG2incombined group. The combined index<1, the two drugs had synergisticeffects. The IC50value concentration of two drug combination was NCTD(38.1mol/L)+EVO (7.6mol/L) in48h.3．Compared with those in single inducer group, NCTD(10μmol/L)combined with EVO(2μmol/L) up-regulated the stagnation rate of G2/M from (36.13±1.63)%with EVO and (10.67±0.89)%with NCTD to(73.42±1.92)%with combined drug.(P<0.05)4. HepG2cells were induced by NCTD（40μmol/L） and/or EVO（8μmol/L）. The apoptosis rate in liver cancer cell treated by two-drugcombination increased significantly from EVO (15.78±3.08)%andNTCD (11.47±1.60)%to EVO+NTCD (21.86±2.70)%.(P<0.05)5. Compared with the control group, the activity of oxygen enhanced indrug treatment group.(P<0.05)6. Western blotting results demonstrated that the expression level ofBax protein went up significantly (P<0.05), meanwhile Bcl-2proteindropped significantly (P<0.05) in EVO or/and NCTD group. Thecombination of two drugs made this change more significant in proteinexpression.(P<0.05)Conclusions1. EVO combined with NCTD showed synergetic effects onanti-proliferative and apoptosis in liver cancer cells.2. EVO combined with NCTD inhibited the growth of HepG2throughdecreasing the proportion of Bax/Bcl-2, arresting the cell cycle in G2/Mphase, and improving the level of intracellular reactive oxygen species.