塞来昔布联合替莫唑胺对胶质瘤U251细胞作用研究

Effect of celecoxib combined with temozolomide on proliferation and apoptosis of glioma cell line U251 cells

目的探讨环氧合酶抑制剂塞来昔布联合抗肿瘤药物替莫唑胺对神经胶质瘤细胞系U251细胞的体外抗瘤效应及其可能机制。方法分别使用塞来昔布、替莫唑胺和两者联合用药干预U251细胞;显微镜下观察细胞形态学变化;以甲基噻唑基四唑比色法检测药物对U251细胞增殖的影响;细胞划痕实验观察药物对U251细胞迁移能力的影响;采用膜联蛋白Ⅴ-碘化丙啶双染色通过流式细胞仪检测药物对肿瘤细胞凋亡的影响;Western Blot法检测药物对U251细胞中Bcl-2和Bax表达的影响。结果药物处理后,U251细胞出现明显的形态学改变,以联合用药组细胞最为显著;当塞来昔布与替莫唑胺联合应用时,显著增强替莫唑胺对U251细胞增殖的抑制效应,抑制细胞迁移,并促进其凋亡;塞来昔布和替莫唑胺联合作用后,U251细胞Bax表达增高、Bcl-2表达降低。结论塞来昔布和替莫唑胺联合应用可以协同抑制肿瘤细胞增殖、迁移,促进细胞凋亡;其促进细胞凋亡的机制可能与Bax表达的上调以及Bcl-2表达的下调有关。

Objective To study the curative effects of celecoxib combined with antitumor drugs on gliomas and its mechanism. Methods The human glioma cell lines U251 cells were cultured and they treated with celecoxib or temozolomide or both the drugs. The cell morphologic change was observed under the optical microscope. The proliferation of U251 cells was determined by methyl thiaszolyl tetrazoliu assay. The migration ability of U251 cells was determined by wound healing test. The apoptosis of U251 cells were evaluated by flow cytometry with PI-Annexin V staining. The expression of Bcl-2 and Bax in U251 cells were detected by western blot. Results The morphologic changes occurred in U251 cells treated with celecoxib or temozolomide or both the drugs, but the morphologic change in U251 cells treated by both the drugs were most significant. The inhibitory effect of celecoxib combined with temozolomide on U251 cells proliferation was significantly stronger than those of celecoxib or temozolomide （P〈0.01）. The apoptosis rate in U251 cells treated by both the drugs was significantly higher than those in U251 cells treated by celecoxib or temozolomide alone （P〈0.01）. The celecoxib combined with temozolomide more significantly inhibited the migration of U251 cells compared to celecoxib or temozolomide alone. The level of Bax expression was significantly higher and the level of Bcl-2 expression was significantly lower in U251 cells treated by both the drugs than those in U251 cells treated by celecoxib or temozolomide alone. Conclusion The combinative application of celecoxib and temozolomide can promote the apoptosis and inhibit the proliferation and migration of U251 cells in a synergistic manner. These effects of celecoxib combined with temozolomide may be related with the upregnlation of Bax expression and down regulation of Bcl-2 expression.