选择性环氧化酶2抑制剂NS-398对胃癌细胞AGS增殖的抑制作用

目的观察选择性环氧化酶2(COX-2)抑制剂NS-398对胃癌细胞AGS增殖的抑制作用,并探讨其相关机制。方法 NS-398 25,50和100μmol·L-1作用于AGS细胞0～48 h,用CCK-8法检测细胞存活率。NS-398 50μmol·L-1作用于AGS细胞48 h,用流式细胞仪检测细胞凋亡率,实时荧光定量PCR检测Notch信号通路相关基因的表达,Western印迹法检测Notch胞内结构域(NICD)及下游靶基因NF-κB和毛蛋白和断裂1增强子(Hes-1)蛋白表达。结果 NS-398 25,50和100μmol·L-1能抑制胃癌细胞AGS增殖,并呈时间(r时间=-1.00,P=0.003,50μmol·L-1)和浓度(r浓度=-0.999,P=0.027,48 h)依赖性。NS-39850μmol·L-1作用48 h,AGS细胞凋亡率为(20.1±3.5)%,比正常对照组(3.5±1.4)%明显增加(P<0.05);Notch信号通路受体Notch1和Notch2及Notch信号通路配体δ样1(DLL1)和锯齿状1(JAG1)mRNA表达无明显变化,Notch下游靶基因Hes-1和NF-κB mRNA表达较正常对照组明显减少(P<0.05);NICD,Hes-1和NF-κB蛋白表达明显减少(P<0.05)。结论选择性COX-2抑制剂NS-398可能通过抑制Notch信号途径抑制胃癌细胞AGS增殖。

Inhibition of cyclooxygenase-2 selective inhibitor NS-398 on proliferation of AGS gastric cancer cells

OBJECTIVE To investigate the inhibition of cyclooxygenase-2 selective inhibitor NS-398 on gastric cancer cell proliferation and the possible mechanism. METHODS After AGS cells were treated with NS-398 25, 50 and 100 μmol·L^-1 for 48 h, AGS cell viability was determined by CCK-8 assay. Flow cytometric analysis was used to detect the apoptosis of AGS cells after 48 h treatment with NS-398 50 μmol·L^-1. The expression of Notch signal pathway related genes was evaluated by real-time PCR. The protein expression of Notch intracellular domain (NICD), NF-κB2 and Hes-1(hairy and enhancer of split 1) in AGS cells was determined by Western blotting. RESULTS Treatment with NS-398 potently induced apoptosis of AGS cells in a time-(r_Time=-1.00,P=0.003, 50 μmol·L^-1) and concentration-(r_{Concentration}=-0.999,P=0.027, 48 h) dependent manner. The apoptosis rate of AGS cells treated with NS-398 50 μmol·L^-1 for 48 h increased to (20.1±3.5)% compared with the control group (3.5±1.4)%(P<0.05). In addition, the mRNA expression of Hes-1 and NF-κB2 was reduced(P<0.05) 24 h after treatment of NS-398 50 μmol·L^-1 while Notch1, Notch2, DLL1(delta like 1) and JAG1(jagged-1) mRNA expression was not changed obviously. Moreover, NICD, Hes-1 and NF-κB2 protein expression was decreased(P<0.05). CONCLUSION The cyclooxygenase-2 selective inhibitor NS-398 reduces proliferation of AGS gastric cancer cells through the Notch signal pathway.