NS-398和DHA联合抑制β-eatenin、c-mye对胆管癌QBC939细胞凋亡的影响  

The influence of NS-398 combined with DHA on the apoptosis of bile duct cancer cells QBC939 through inhibiting β-catenin and c-myc

作　　者：姜海涛[1] 张顺[2] 任燕萍[3] 史佩娟 陈云杰[1] Jiang Haitao;Zhang Shun;Ren Yanping;Shi Peijuan;Chen Yunjie(Department of General Surgery, Ningbo No. 2 Hospital, Ningbo 315010, China)

机构地区：[1]宁波市第二医院外二科,315010 [2]宁波市第二医院干细胞实验室科,315010 [3]宁波市第二医院药学部,315010 [4]宁波市第二医院财务科,315010

出　　处：《中华普通外科杂志》2018年第5期416-419,共4页Chinese Journal of General Surgery

基　　金：华美研究基金资助项目（2017HMKY11）

摘　　要：目的研究选择性环氧合酶-2（cyclooxygenase-2，COX-2）抑制剂NS-398和二十二碳六烯酸（docosahexaenoic acid，DHA）联合对胆管癌细胞QBC939凋亡的影响及机制。方法体外培养胆管癌QBC939细胞，设立NS-398组、DHA组、联合组和空白组，将0，25，50，100，150，200μmol／L的NS-398和0，15，30，45，60，75μg／ml的DHA分别联合作用于QBC939细胞；应用CCK8法检测QBC939细胞的生长抑制率：采用流式细胞术检测细胞凋亡：应用实时荧光定量PCR、Western blot和ELISA检测细胞中β-catenin和c-myc的表达。结果NS-398和DHA在体外均能抑制QBC939细胞生长，NS-398为100pomol／L联合DHA为45μg／ml作用后，细胞生长抑制率达90％（F=5．85，P〈0．05），二者联合能明显促进QBC939细胞凋亡（F=8．16，P〈0．01）。实时荧光定量PCR、Western blot和ELISA实验均显示联合能明显抑制β-catenin（F=7．61，P〈0．01）、（F=7．75，P〈0．01）、（F=8．17，P〈0．01）和c-myc（F=7．92，P〈0．01）、（F=8．76，P〈0．01）、（F=8．12，P〈0．01）表达。结论NS-398和DHA联合能明显抑制QBC939细胞生长，促进早期凋亡，二者联合对QBC939细胞生长的抑制作用可能是通过抑制β-catenin和c-myc的表达，促进细胞早期凋亡。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 μg/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 13-catenin （F =7.75 ,P 〈 0.01）, （F=8.17,P〈0.01） andc-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.

关 键 词：胆管肿瘤 细胞凋亡 Β-CATENIN C-MYC 

分 类 号：R735.8[医药卫生—肿瘤]