Inhibitory Effect And Mechanism Of EVO Combined With GEM On Human Tongue Squamous Cell Carcinoma In Vitro

Objective: In this study,we investigate whether Evodiamine combined with chemotherapy drugs have synergistic inhibition effect on tongue squamous carcinoma,and its possible mechanism.Materials and Methods: The influence of Evodiamine combined with Gemcitabine in different sequence and dosing ratio on tongue squamous cancer cell proliferation is determined by MTT assay and drug interactions is analyzed by the Chou-Talalay.The cancer cell clone ability,apoptosis and migration is determined by plate clone formation assay,Annexin V/PI staining,Hoechst33342 staining and wound healing test.The NF-?B p65 activity and its downstream protein is determined by western blot in order to determine whether the activity of NF-?B signal involved in Evodiamine combined with gemcitabine for produce synergistic.Results: 1.The sequential administration of Evodiamine followed by gemcitabine meanwhile the dosing ratio of E: G=2:1 may be the optimal schedule of this combination.2.Combined administration on cells can significantly inhibit the clone formation ability(P < 0.05)and the migration ability,and increase the apoptosis compared with the control group and single drug group.3.Evodiamine can significantly inhibit NF-?B p65 expression in CAL-27 cell line,but there are no differences in Tca8113 cell line.Cancer cells express NF-?B p65 itself and cells exposured to Gemcitabine express NF-?B p65 more than former,but there are no statistically significant difference between both.Combined administration can reduce the NF-?B p65 expression,Bcl-2 and Bcl-xl.Conclusions:1.different sequence and different dosing ratio administration of Evodiamine and Gemcitabine can affect the interaction between antitumor drugs.2.Evodiamine may inhibit cell proliferation and induce apoptosis by reducing the NF-?B signaling afterwards reducing the expression of Bcl-2 and Bcl-xl.Combined administration can enhance the cytotoxicity of chemical drugs on tongue squamous cancer cells,depress the clone formation and migration ability.