顺铂联合COX-2选择性抑制剂NS-398对食管癌Eca-109细胞增殖和凋亡的影响

Effect of combined cisplatin and COX-2 selective inhibitor NS-398 on proliferation and apoptosis of human esophageal cancer Eca-109 cells

目的体外观察顺铂(diamminedichloroplatinum,DDP)以及DDP联合环氧合酶-2(cyclooxygenase,COX-2)选择性抑制剂NS-398对人食管癌Eca-109细胞增殖和凋亡的影响,比较单独应用DDP与DDP联合NS-398应用于食管癌Eca-109细胞的抗肿瘤效应。方法 CCK-8法检测不同浓度的DDP(1.25、2.50、5.00、10.00、20.00mg/L)单独作用及DDP联合NS-398(1.00、10.00μmol/L)作用Eca-109细胞24h的细胞增殖抑制率。琼脂糖凝胶电泳技术检测DDP(1.25、2.50、5.00mg/L)单用及联合NS-398(1.00、10.00、80.00μmol/L)作用Eca-109细胞24h和48h的DNA Ladder出现情况。透射电镜观察DDP单用以及联合NS-398作用于Eca-109细胞24h超微结构的变化。药物未处理组作为对照组。结果细胞增殖抑制实验表明顺铂、NS-398对Ec-109细胞有剂量依赖性抑制增殖作用(P<0.05)。顺铂联合NS-398用药组细胞增殖抑制率明显高于同浓度顺铂单用组,且随添加的NS-398的剂量的增加而增加,DDP联合NS-398用药组对Ec-109细胞的增殖抑制效应和诱导凋亡效应明显高于DDP单药组,二者具有协同效应。结论与DDP单用相比,添加低剂量NS-398能够增强DDP对Eca-109细胞的增殖抑制效应和诱导凋亡效应。

Objective To observe the effect of diamminedichloroplatinum（DDP）and combined DDP and cyclooxygenase（COX-2）selective inhibitor NS-398 on proliferation and apoptosis of esophageal cancer Eca-109 cells in vitro and their anti-tumor effect.Methods CCK-8 assay was used to detect the inhibitory rate of DDP at the concentrations of 1.25,2.5,5,10,and 20 mg/L and combined DDP and NS-398 at the concentrations of 1.0 and 10.0 μmol/L for human esophageal cancer Eca-109 cells 24 h after they were used.Agarose gel electrophoresis was performed to show the DNA ladder of human esophageal Eca-109cancer cells in 24 h and 48 h after they were treated with DDP at the concentration of 1.25,2.5,and 5.0 mg/L,and combined DDP and NS-398 at the concentration of 1.0,10.0,and 80.0 mg/L.Transmission electron microscopy was employed to observe the ultra-structure of Eca-109 cells 24 h after they were treated with DDP and combined DDP and NS-398,respectively.Non-drug treated esophageal cancer Eca-109 cells served as a control group.Results Cell growth inhibition test showed that DDP and NS-398 inhibited the proliferation of esophageal cancer Eca-109 cells in a dose-dependent manner（P0.05）.The growth inhibition rate and apoptosis rate of combined DDP and NS-398 were significantly higher than those of DDP for esophageal cancer Eca-109 cells,indicating that DDP and combined DDP and NS-398 exhibited a synergistic effect on the proliferation and apoptosis of esophageal cancer Eca-109 cells.Conclusion The effect of DDP in combination with a low dose of NS-398 is stronger than that of DDP on the proliferation and apoptosis of esophageal cancer Eca-109 cells.