The effect of docosahexaenoic acid combined with cyclooxygenase-2 selective inhibitor NS-398 on the apoptosis of bile duct cancer cells QBC939
10.3760/cma.j.issn.1007-8118.2018.05.013
- VernacularTitle:二十二碳六烯酸联合选择性环氧合酶-2抑制剂NS-398对人胆管癌QBC939细胞凋亡的影响
- Author:
Haitao JIANG
1
;
Wei ZHOU
;
Lian'gang MAO
;
Yunjie CHEN
;
Jingyu CAO
Author Information
1. 315010,宁波市第二医院外二科
- Keywords:
Docosahexaenoic acid (DHA);
NS-398;
Cholangiocarcinoma;
Apoptosis;
β-catenin;
Cyclooxygenase-2 (COX-2)
- From:
Chinese Journal of Hepatobiliary Surgery
2018;24(5):336-340
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of docosahexaenoic acid (DHA) combined with cyclooxygenase-2 (COX-2) selective inhibitor NS-398 on the apoptosis of cholangiocarcinoma QBC939 cells and the mechanism.Methods In vitro,cholangiocarcinoma QBC939 cells were treated with 0,15,30,45,60 and 75 μg/ml DHA with 0,25,50,100,150 and 200 μmol/L NS-398,respectively.The absorbances of the QBC939 cells were measured by CCK8 and its growth inhibition ratios were analyzed.Flow cytometry was applied to detect cell apoptosis.The level of β-catenin and COX-2 mRNA and protein were measured by real-time PCR,immunocytochemistry and enzyme-linked immunoadsordent assay,respectively.Results DHA combined with NS-398 could significantly suppress the growth of QBC939 cells (P ＜ 0.05).When the concentration of DHA went up to 45 μg/ml and NS-398 was 100 μmol/L,the relative growth inhibition rate of QBC939 cells was 90.0％.If the concentrations were increased,the result showed no significant differences.Furthermore,flow cytometry analysis indicated that DHA combined with NS-398 could induce QBC939 cells apoptosis at the early stage,and the apoptosis rate was significantly different between the experimental and control groups (P ＜ 0.01).Real-time PCR showed low β-catenin and COX-2 expression in QBC939 cells disposed by DHA combined with NS-398,and their expression were significantly different between the experimental and control groups (P ＜ 0.01).Immunocytochemistry and ELISA demonstrated that DHA combined with NS-398 could decrease β-catenin and COX-2 protein expression in QBC939 cells.Conclusion DHA combined with NS-398 induced apoptosis and inhibited proliferation of cholangiocarcinoma cells QBC939 in vitro through targeting β-catenin and COX-2.
