A Study Of Effect Of Sinapine Thiocyanate’s On The Proliferation,migration And Invation,promoting The Sensitivity Of Hepatocellular Carcinoma Cell To Drug

Objective:This study detectd the effect of sinapine thiocyanate on influencing the proliferation,migration and invasion of human hepatocellular carcinoma cell and promoting the sensitivity of human hepatocellular carcinoma cell to gemcitabine,as well as the molecular mechanism.Methods: Firstly,the effect of sinosine thiocyanate on the proliferation,migration and invasion of human hepatocellular carcinoma cells and its effect on enhancing the sensitivity of hepatocellular carcinoma cells to gemcitabine were investigated.CCK-8,scratch assay and Transwell assay were used respectively to detect the effects of different concentrations(0,25,50 and 100 μm/L)of sinosine thiocyanate on the proliferation,migration and invasion ability of SMMC7721 cells.CCK-8,colony formation assay and flow cytometry were used repectively to detect the effects of sinapine thiocyanate combined with gemcitabine on the proliferation,colony formation and apoptosis of Hep G2 cells.Rhodamine 123 was used to detect the effect of sinapine thiocyanate on the drug efflux ability of Hep G2cells;then the mechanism of sinosine thiocyanate was discussed.Computer molecular docking technique was used to verify the binding ability of sinosine thiocyanate to protein targets PTGS1,PTGS2,and the binding ability of sinapine thiocyanate to bind with multi-drug resistance related proteins ABCB1 and ABCG2;Western blot was used to detect the effects of sinosine thiocyanate on the protein expression levels of PTGS1,PTGS2,Bcl2,Bax,MMP2 and MMP9 in SMMC-7721 cells and the effect of sinapine thiocyanate combined with gemcitabine on the expression of ABCB1,ABCG2,caspase-8,Cleaved caspase-8,Bcl2 and Bax in Hep G2 cells.Results:Sinosine thiocyanate can inhibite the proliferation,migration and invasion of hepatocellular carcinoma cells SMMC 7721.Sinosine thiocyanate can bind with PTGS1 and PTGS2 to decrease the protein expressions of PTGS1,PTGS2,Bcl2,MMP2 and MMP9 in HCC SMMC-7721 cells(P<0.05),increasing the protein expression of Bax(P<0.05).Sinapine thiocyanate can enhance the inhibitory effects of gemcitabine on Hep G2 cell proliferation and colony formation(P<0.05)and the ability of gemcitabine to induce the apoptosis in Hep G2(P<0.05);Sinapine thiocyanate can stably bind with multi-drug resistance related proteins ABCB1 and ABCG2 and reduced the drug efflux ability of Hep G2 cells,by inhibiting the expression of ABCB1 and ABCG2 proteins,as well as enhancing the reduction the expression of Bcl2 and the increased expression of cleaved caspase-8 and Bax induced by gemcitabine(P<0.05).Conclusions:Sinapine thiocyanate can inhibit the proliferation、migration and invasion of hepatocellular carcinoma cells SMMC 7721 via targeting PTGS1 and PTGS2,promoting the sensitivity of human hepatocellular carcinoma cell Hep G2 to gemcitabine via targeting ABCB1 and ABCG2.