三氧化二砷联合塞来昔布对卵巢癌细胞的增殖抑制和凋亡诱导作用

Combination of As_2O_3 and celecoxib can inhibit the proliferation and induce apoptosis of ovarian cancer cells

目的 :探讨在上皮性卵巢癌中三氧化二砷(As2O3)与环氧合酶-2(cyclooxygenase-2,Cox-2)抑制剂塞来昔布(celecoxib)联合用药的有效性及其作用机制。方法 :以上皮性卵巢癌细胞株OVCAR-3为研究对象,将As2O3和celecoxib单药及两药联合作用该细胞株48 h后,采用MTT体外药敏实验检测细胞增殖抑制作用,并用金正均q值法计算联合用药的q值。然后采用Hoechest33258凋亡染色和FCM法检测细胞凋亡及细胞周期变化,蛋白质印迹法检测Bcl-2、Bax和切割型caspase-3等凋亡相关蛋白的表达水平变化。结果 :As2O3和celecoxib对OVCAR-3细胞均具有剂量依赖性的生长抑制作用,两种药物单独作用的半数抑制浓度(50%concentration of inhibition,IC50)值分别为4.72和42.58μmol/L。两药联合作用后,OVCAR-3细胞的增殖抑制率和凋亡率均大于任一单药作用组(P<0.05),联合用药的q值均大于1.15。As2O3和celecoxib联合用药后OVCAR-3细胞发生明显的G2/M期阻滞,而且Bcl-2表达明显下调,Bax和caspase-3表达则明显上调(均P<0.05)。结论 :As2O3和Cox-2抑制剂celecoxib单药作用均对上皮性卵巢癌细胞的生长有抑制作用,两者联合用药后则具有显著的协同抗肿瘤效应。推测As2O3联合celecoxib是一个具有潜在研究价值的新的化疗方案。

Objective： To investigate the effectiveness of arsenic trioxide （As2O3） in combination with celecoxib, an inhibitor of cyclooxygenase-2 （Cox-2）, for epithelial ovarian cancer cells, and its mechanism. Methods： The epithelial ovarian cancer OVCAR-3 cells were treated with As2O3 and celecoxib alone and in combination for 48 h. Then, the cell proliferation was examined by MTT assay. The interaction of As2O3 and celecoxib was estimated by JIN Zhengjun’s method, and the q value was calculated. The apoptosis and cell cycle distribution were detected by Hoechest33258 fluorescence staining and flow cytometry. The expressions of Bcl-2, Bax and cleaved caspase-3 were detected by Western blotting. Results： Dose-dependent anti-proliferation effects on OVCAR-3 cells were observed when As2O3 and celecoxib alone were administered, and the corresponding half inhibitory concentration （IC50） values were 4.72 and 42.58 μmol/L, respectively. When these two agents were applied simultaneously, the growth inhibition rate and the apoptosis rate were significantly higher than those of the OVCAR-3 cells treated with As2O3 and celecoxib alone （both P 〈 0.05）, and the q value was greater than 1.15. After treatment with As2O3 combined with celecoxib, the OVCAR-3 cells were significantly blocked at G2/M phase （P 〈 0.05）; while the expression of Bcl-2 was declined obviously, but the expressions of Bax and cleaved caspase-3 were markedly increased （all P 〈 0.05）. Conclusion： As2O3 and celecoxib alone can exert inhibitory effect on the growth of epithelial ovarian cancer cells. There is a synergistic effect when As2O3 and celecoxib were used in combination, so that the combination of As2O3 and celecoxib is predicted to be a potentially promising chemotherapeutic regimen for ovarian cancer.