Salizzato Valentina, Borgo Christian, Cesaro Luca, Pinna Lorenzo A, Donella-Deana Arianna
Department of Biomedical Sciences and CNR Institute of NeuroSciences, University of Padova, 35131 Padova, Italy.
Oncotarget. 2016 Apr 5;7(14):18204-18. doi: 10.18632/oncotarget.7569.
Chronic myeloid leukaemia (CML) is a myeloproliferative disorder promoted by the constitutive tyrosine kinase activity of Bcr-Abl oncoprotein. Although treatment with the Bcr-Abl-inhibitor imatinib represents the first-line therapy against CML, almost 20-30% of patients develop chemotherapeutic resistance and require alternative therapy. Here we show that a strong hyper-phosphorylation/activation of ERK1/2, Akt Ser473, and 40S ribosomal protein S6 (rpS6) is detectable in imatinib-resistant KCL22 and K562 CML cells as compared to the -sensitive cell variants. In imatinib-resistant CML cells, high concentration of imatinib is required to strongly inhibit Bcr-Abl, ERK1/2 and Akt Ser473 phosphorylation, but under these conditions the phosphorylation of rpS6, a common downstream effector of MEK/ERK1/2 and PI3K/Akt/mTOR pathways is only slightly reduced. By contrast, down-regulation of the protein kinase CK2 by the inhibitor CX-5011 or by silencing the CK2 subunits does not affect the activation state of MEK/ERK1/2 or PI3K/Akt/mTOR signalling, but causes a drop in rpS6 phosphorylation in parallel with reduced protein synthesis. CK2-inhibition by CX-5011 induces cell death by apoptosis and acts synergistically with imatinib or the MEK-inhibitor U0126 in reducing the viability of imatinib-resistant CML cells. The ternary mixture containing CX-5011, imatinib and U0126 represents the most effective synergistic combination to counteract CML cell viability. These results disclose a novel CK2-mediated mechanism of acquired imatinib-resistance resulting in hyper-phosphorylation of rpS6. We suggest that co-targeting CK2 and MEK protein kinases is a promising strategy to restore responsiveness of resistant CML cells to imatinib.
慢性髓性白血病(CML)是一种由Bcr-Abl癌蛋白的组成型酪氨酸激酶活性引发的骨髓增殖性疾病。尽管使用Bcr-Abl抑制剂伊马替尼进行治疗是针对CML的一线疗法,但几乎20%-30%的患者会产生化疗耐药性,需要替代疗法。在此我们表明,与敏感细胞变体相比,在伊马替尼耐药的KCL22和K562 CML细胞中可检测到ERK1/2、Akt Ser473和40S核糖体蛋白S6(rpS6)的强烈过度磷酸化/激活。在伊马替尼耐药的CML细胞中,需要高浓度的伊马替尼才能强烈抑制Bcr-Abl、ERK1/2和Akt Ser473的磷酸化,但在这些条件下,rpS6的磷酸化(MEK/ERK1/2和PI3K/Akt/mTOR途径的常见下游效应物)仅略有降低。相比之下,抑制剂CX-5011下调蛋白激酶CK2或沉默CK2亚基并不影响MEK/ERK1/2或PI3K/Akt/mTOR信号的激活状态,但会导致rpS6磷酸化下降,同时蛋白质合成减少。CX-5011抑制CK2通过凋亡诱导细胞死亡,并与伊马替尼或MEK抑制剂U0126协同作用,降低伊马替尼耐药CML细胞的活力。包含CX-5011、伊马替尼和U0126的三元混合物是对抗CML细胞活力最有效的协同组合。这些结果揭示了一种新的CK2介导的获得性伊马替尼耐药机制,导致rpS6过度磷酸化。我们认为,共同靶向CK2和MEK蛋白激酶是恢复耐药CML细胞对伊马替尼反应性的一种有前景的策略。