Division of Hematology, Department of Biomedical Sciences, Ospedale Ferrarotto, University of Catania, Catania, Italy.
Eur J Haematol. 2011 Mar;86(3):216-25. doi: 10.1111/j.1600-0609.2010.01570.x. Epub 2011 Feb 8.
BCR/ABL positive cells are known to be resistant to DNA damage induced by chemotherapy while they are sensitive to imatinib (IM), a tyrosine kinase inhibitor (TKI). To evaluate whether this drug can increase the activity of cytotoxic drugs on BCR/ABL positive cells, we measured the toxicity of cytosine arabinoside (ARA-C), hydroxyurea (HU) and melphalan (MEL), after a pretreatment of 24 h with IM on K562 cell line. The highest cytotoxic effect was seen when the TKI was followed by MEL; our results indicate that inhibition of BCR/ABL activity by IM increased the cytotoxicity of MEL by favoring the DNA damage induced by this drug and by shortening the time for DNA repair at the G2/M checkpoint. A stronger activation of some genes involved in both intrinsic and extrinsic apoptotic pathways was also observed with IM/MEL combination compared to IM or MEL alone. The drugs association was further tested in a type of BaF3 cells (TonB.210) where the BCR-ABL expression is inducible by doxycycline; in this model it was confirmed that a reduction of BCR/ABL activity resulted in an increased susceptibility to the cytotoxic effect of MEL. Furthermore, we studied the effect of IM/MEL treatment on the proliferative potential of myeloid progenitors of six CML patients at diagnosis. The analysis of CFU-GM and BFU-E colonies demonstrated that the IM/MEL combination was more effective than IM alone in reducing the overall number of colonies and the number of copies of BCR/ABL. In conclusion, our work shows that inhibition of BCR/ABL activity increases the toxicity of MEL and allows an efficient killing of leukemic cells, suggesting that a clinical development of this approach could have therapeutic advantages for CML patients.
已知 BCR/ABL 阳性细胞对化疗诱导的 DNA 损伤具有抗性,而对伊马替尼(IM),一种酪氨酸激酶抑制剂(TKI)敏感。为了评估这种药物是否能增加细胞毒性药物对 BCR/ABL 阳性细胞的活性,我们在 K562 细胞系上用 IM 预处理 24 小时后,测量了胞嘧啶阿拉伯糖苷(ARA-C)、羟基脲(HU)和美法仑(MEL)的毒性。当 TKI 后接 MEL 时,观察到最高的细胞毒性作用;我们的结果表明,通过抑制 BCR/ABL 活性,IM 增加了 MEL 的细胞毒性,有利于该药物诱导的 DNA 损伤,并缩短了 G2/M 检查点的 DNA 修复时间。与 IM 或 MEL 单独使用相比,还观察到 IM/MEL 联合使用时一些参与内在和外在凋亡途径的基因的激活更强。在一种 BaF3 细胞(TonB.210)中进一步测试了药物的联合使用,这种细胞中 BCR-ABL 的表达可以通过强力霉素诱导;在该模型中证实,BCR/ABL 活性的降低导致对 MEL 的细胞毒性作用的敏感性增加。此外,我们研究了 IM/MEL 治疗对 6 例 CML 患者诊断时髓样祖细胞增殖潜力的影响。CFU-GM 和 BFU-E 集落的分析表明,与 IM 单独治疗相比,IM/MEL 联合治疗更有效地减少集落总数和 BCR/ABL 的拷贝数。总之,我们的工作表明,抑制 BCR/ABL 活性增加了 MEL 的毒性,并能有效地杀死白血病细胞,这表明这种方法的临床发展可能对 CML 患者具有治疗优势。