松油烯-4-醇对人多发性骨髓瘤U266细胞增殖及凋亡的影响

Effect of Terpinen-4-ol on Proliferation and Apoptosis of Human Multiple Myeloma Cells U266

目的探讨松油烯-4-醇（T4O）对人多发性骨髓瘤U266细胞增殖及凋亡的影响及其作用机制。方法不同浓度（5~80μmol/L）T4O体外作用于U266细胞。CCK-8法检测增殖抑制率;AO/EB染色,倒置显微镜观察细胞凋亡形态;比色法检测Caspase-3酶活性;流式细胞术检测细胞凋亡及周期;JC-1染色检测细胞线粒体膜电位。结果 T4O对U266细胞有增殖抑制作用,呈时间-剂量依赖性（P〈0.05）。T4O诱导U266细胞凋亡,呈剂量依赖性（P〈0.05）,随药物浓度增加,Caspase-3酶活性逐渐增强（P〈0.05）,线粒体膜电位逐渐降低（P〈0.05）,细胞周期被阻滞在G2期。结论 T4O对U266细胞有抑制增殖及促凋亡作用,其机制是激活细胞凋亡途径关键酶Caspase-3,使细胞线粒体膜电位降低,并且使细胞周期阻滞在G2期,进而抑制肿瘤细胞分化与增殖。

Objective To investigate the effect of Terpinen-4-ol（T4O） on the apoptosis and proliferation of human multiple myeloma cells U266 and related mechanism. Methods U266 cells were incubated with different concentrations of T4O（5-80μmol/L） in vitro. Then cell proliferation inhibition rate was determined by cell counting kits（CCK-8） assay. The change of apoptosis morphology was observed by inverted microscope after AO/EB staining. Caspase-3 activities were measured with a colorimetric method. Apoptosis and cell cycles were analyzed by FCM, and mitochondrial membrane potential（MMP） were detected by JC-1 staining. Results T4 O exhibited obvious inhibition effect on the proliferation of U266 cells in a timedose-dependent manner（P0.05）. Meanwhile, T4 O induced the apoptosis of U266 cells in a dose-dependent manner（P0.05）. The activity of Caspase-3 was enhanced in a dose-dependent manner（P0.05）. MMP were reduced in a dose-dependent manner（P0.05）. Cell cycles were arrested in G2 phase. Conclusion T4 O exhibits markedly effect of proliferation inhibition and apoptosis-inducing on U266 cells by activating the key enzyme Caspase-3 of apoptosis pathway. MMP are declined in a dose-dependent manner and cell cycles are arrested in G2 phase as well, thus, to inhibit the proliferation and differentiation of tumors.