松油烯-4-醇对人多发性骨髓瘤U266细胞增殖及凋亡的影响

涂云华; 李明娥; 康颖倩; 荣冬芸; 周英; 薛月萃; 叶振源; 昝雪娟; 曹煜

doi:10.3971/j.issn.1000-8578.2016.05.008

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

摘要

摘要:
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（5~80 μmol/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。

Abstract:
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 T4O exhibited obvious inhibition effect on the proliferation of U266 cells in a time-dose-dependent manner(P < 0.05). Meanwhile, T4O induced the apoptosis of U266 cells in a dose-dependent manner(P < 0.05). The activity of Caspase-3 was enhanced in a dose-dependent manner(P < 0.05). MMP were reduced in a dose-dependent manner(P < 0.05). Cell cycles were arrested in G₂ phase.

Conclusion T4O 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.

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