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目的观察环氧化酶-2(COX-2)抑制剂塞来昔布对骨髓瘤细胞株U266细胞的增殖抑制和凋亡诱导的影响,并通过比较单用塞来昔布和加用Caspase-3阻断剂Ac-DEVD-FMK后的U266细胞株的凋亡率以及Caspase-3的表达,探讨其诱导U266细胞株凋亡的分子机制。方法骨髓瘤细胞株U266细胞经不同浓度的塞来昔布处理后,应用CCK-8体外药物筛选法测定受试样品塞来昔布对人骨髓瘤细胞U266体外增殖有无抑制作用及作用强弱;Annexin V-FITC染色流式细胞术检测细胞凋亡;Western blot法测定不同实验组样品中Caspase-3总量的表达。结果塞来昔布浓度>40μmol/L能抑制人骨髓瘤细胞U266体外增值,并呈浓度依赖性,IC50=66.83μmol/L。塞来昔布在40~120μmol/L浓度范围内能诱导人骨髓瘤细胞U266细胞凋亡,呈浓度依赖性。用Ac-DEVD-FMK阻断Caspase-3活性,塞来昔布诱导的U266细胞凋亡明显受抑。结论塞来昔布可诱导骨髓瘤细胞株U266细胞株凋亡,其机制涉及Caspase-3依赖性凋亡调节信号通路。
Objective To observe the effect of cyclooxygenase-2 (COX-2) inhibitor celecoxib on the proliferation and apoptosis induction of myeloma cell line U266. By comparing with celecoxib alone and adding Caspase- 3 inhibitor Ac-DEVD-FMK after U266 cell line apoptosis and Caspase-3 expression, to explore the molecular mechanism of its induction U266 cell apoptosis. Methods After myeloma cell line U266 cells were treated with different concentrations of celecoxib, CCK-8 in vitro drug screening method was used to determine whether celecoxib inhibited the proliferation of human myeloma cells U266 in vitro and its effect was strong Annexin V-FITC staining flow cytometry was used to detect apoptosis; Western blot was used to determine the total amount of Caspase-3 in different experimental groups. Results Celecoxib concentration> 40μmol / L inhibited the proliferation of human myeloma cell line U266 in a concentration-dependent manner with IC50 of 66.83μmol / L. Celecoxib could induce the apoptosis of human myeloma cell line U266 in a dose-dependent manner in the concentration range of 40 ~ 120μmol / L. Activation of Caspase-3 by Ac-DEVD-FMK blocked the apoptosis of U266 cells induced by celecoxib. Conclusion Celecoxib can induce the apoptosis of myeloma cell line U266, which is involved in the Caspase-3-dependent apoptosis-regulating signaling pathway.