Objective: To investigate the toxic effect of chlorambucil combined with ibrutinib on mantle cell lymphoma (MCL) cell line Jeko-1 and its related mechanism.
Methods: The MCL cell line Jeko-1 was incubated with different concentrations of chlorambucil or ibrutinib or the combination of the two drugs, respectively. CCK-8 assay was used to detect the proliferation of the cells, and Western blot was used to measure the protein expression levels of BCL-2, caspase-3, PI3K, AKT and P-AKT.
Results: After Jeko-1 cells were treated with chlorambucil (3.125, 6.25, 12.5, 25, 50 μmol/L) and ibrutinib (3.125, 6.25, 12.5, 25, 50 μmol /L) alone for 24, 48, 72h respectively, the cell proliferation was inhibited in a time- and dose-dependent manner. Moreover, the two drugs were applied in combination at low doses (single drug inhibition rate<50%), and the results showed that the combination of two drugs had a more significant inhibitory effect (all P < 0.05). Compared with the control group, the apoptosis rate of the single drug group of chlorambucil (3.125, 6.25, 12.5, 25, 50 μmol/L) and ibutinib (3.125, 6.25, 12.5, 25, 50 μmol/L) was increased in a dose-dependent manner. The combination of the two drugs at low concentrations (3.125, 6.25, 12.5 μmol/L) could significantly increase the apoptosis rate compared with the corresponding concentration of single drug groups (all P < 0.05). Compared with control group, the protein expression levels of caspase-3 in Jeko-1 cells were upregulated, while the protein expression levels of BCL-2, PI3K, and p-AKT/AKT were downregulated after treatment with chlorambucil or ibrutinib alone. The combination of the two drugs could produce a synergistic effect on the expressions of the above-mentioned proteins, and the differences between the combination group and the single drug groups were statistically significant (all P < 0.05).
Conclusion: Chlorambucil and ibrutinib can promote the apoptosis of MCL cell line Jeko-1, and combined application of the two drugs shows a synergistic effect, the mechanism may be associated with the AKT-related signaling pathways.
题目: 苯丁酸氮芥联合伊布替尼对套细胞淋巴瘤Jeko-1细胞株的作用及相关机制.
目的: 探讨苯丁酸氮芥联合伊布替尼对套细胞淋巴瘤Jeko-1细胞株的抑制作用及相关机制。.
方法: 分别将不同浓度的苯丁酸氮芥、伊布替尼单药及两药联合作用于Jeko-1细胞株，采用CCK-8检测细胞增殖情况，同时采用Western blot检测细胞中BCL-2、caspase-3、PI3K、AKT、P-AKT蛋白表达的变化。.
结果: 分别用苯丁酸氮芥（3.125、6.25、12.5、25、50 μmol/L）及伊布替尼（3.125、6.25、12.5、25、50 μmol/L）单药处理Jeko-1细胞株24、48和72 h后，细胞增殖均受到抑制，且抑制作用呈时间及剂量依赖性，两药联合时抑制作用更明显（均P <0.05）。与对照组相比，苯丁酸氮芥（3.125、6.25、12.5、25、50 μmol/L）及伊布替尼（3.125、6.25、12.5、25、50 μmol/L）单药组细胞凋亡率增加，呈剂量依赖性；与相应剂量（3.125、6.25、12.5 μmol/L）单药组比较，两药联合应用组Jeko-1细胞株的凋亡率明显增加（均P <0.05）。与单药组相比，两药联合时可产生协同作用，Jeko-1细胞的caspase-3表达明显增强，BCL-2、PI3K、p-AKT/AKT表达明显减弱（均P <0.05）。.
结论: 苯丁酸氮芥及伊布替尼能够促进套细胞淋巴瘤Jeko-1细胞株的凋亡，两药联合应用可以产生协同作用，其主要是通过AKT相关信号通路发挥作用。.
Keywords: cell apoptosis; chlorambucil; ibrutinib; mantle cell lymphoma.