The influence of NS-398 combined with DHA on the apoptosis of bile duct cancer cells QBC939 through inhibiting β-catenin and c-myc / 中华普通外科杂志

ABSTRACT

Objective To investigate the effect of NS-398 combined with docosahexaenoic acid (DHA) on apoptosis of cholangiocarcinoma QBC939 cells and its mechanism.Methods In vitro,cultured cholangiocarcinoma QBC939 cells were exposed to 0,25,50,100,150 and 200 μmol/L NS-398 with 0,15,30,45,60 and 75 μg/ml DHA respectively.The absorbance of the QBC939 cells were measured by CCK8 and its growth inhibition ratio were calculated.Flow cytometry was applied to detect cell apoptosis.The level of β-catenin and c-myc mRNA and protein were measured by real-time PCR,Western blot and enzyme-linked immunoabsordent assay respectively.Results Exposure to NS-398 combined with DHA suppressed the growth of QBC939 cells.When NS-398 was at 100 μmol/L and DHA at 45 pμg/ml,the relative growth inhibition rate of QBC939 cells was 90％ (F =5.85,P ＜ 0.05).NS-398 combined with DHA promoted QBC939 cells apoptosis at the early stage (F =8.16,P ＜ 0.01).Real-time PCR could detect low β-catenin and c-myc expression in QBC939 cells disposed by NS-398 combined with DHA (F =7.61,P ＜0.01),(F =7.92,P ＜0.01).NS-398 combined with DHA decreased β-catenin (F =7.75,P ＜ 0.01),(F=8.17,P＜0.01) and c-myc (F=8.76,P＜0.01),(F=8.12,P＜0.01) protein expression in QBC939 cells.Conclusion NS-398 combined with DHA promoted apoptosis and inhibit proliferation of cholangiocarcinoma cells QBC939 in vitro possibly through downregulated mRNA and protein expression of β-catenin and c-myc.