Kurosu Tetsuya, Ohki Manabu, Wu Nan, Kagechika Hiroyuki, Miura Osamu
Department of Hematology, Graduate School of Medicine, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo, Japan.
Cancer Res. 2009 May 1;69(9):3927-36. doi: 10.1158/0008-5472.CAN-08-2978. Epub 2009 Apr 14.
Although the BCR/ABL tyrosine kinase inhibitor imatinib is highly effective for treatment of chronic myelogenous leukemia and Philadelphia chromosome-positive (Ph+) acute lymphoblastic leukemia, relapse with emerging imatinib resistance mutations in the BCR/ABL kinase domain poses a significant problem. Here, we show that the multikinase inhibitor sorafenib inhibits proliferation and induces apoptosis at much lower concentrations in Ton.B210 cells when driven by inducibly expressed BCR/ABL than when driven by interleukin-3. The increased sensitivity to sorafenib was also observed in cells inducibly expressing BCR/ABL with the imatinib-resistant E255K or T315I mutation. Sorafenib-induced apoptosis in these cells and Ph+ leukemic cells was synergistically enhanced by rottlerin, bortezomib, or ABT-737 and inhibited by the pan-caspase inhibitor BOC-d-fmk or the overexpression of Bcl-XL. It was further revealed that sorafenib activates Bax and caspase-3 and reduces mitochondrial membrane potential specifically in BCR/ABL-driven cells. Sorafenib also inhibited BCR/ABL-induced tyrosine phosphorylation of its cellular substrates and its autophosphorylation in Ton.B210. It was finally shown that sorafenib inhibits the kinase activity of BCR/ABL as well as its E255K and T315I mutants in in vitro kinase assays. These results indicate that sorafenib induces apoptosis of BCR/ABL-expressing cells, at least partly, by inhibiting BCR/ABL to activate the mitochondria-mediated apoptotic pathway. Thus, sorafenib may provide an effective therapeutic measure to treat Ph+ leukemias, particularly those expressing the T315I mutant, which is totally resistant to imatinib and the second generation BCR/ABL inhibitors.
尽管BCR/ABL酪氨酸激酶抑制剂伊马替尼对慢性粒细胞白血病和费城染色体阳性(Ph+)急性淋巴细胞白血病的治疗非常有效,但BCR/ABL激酶结构域中出现的伊马替尼耐药突变导致的复发仍是一个重大问题。在此,我们表明,在诱导表达BCR/ABL的Ton.B210细胞中,多激酶抑制剂索拉非尼在比由白细胞介素-3驱动时低得多的浓度下就能抑制增殖并诱导凋亡。在诱导表达具有伊马替尼耐药性E255K或T315I突变的BCR/ABL的细胞中也观察到对索拉非尼的敏感性增加。在这些细胞和Ph+白血病细胞中,索拉非尼诱导的凋亡被rottlerin、硼替佐米或ABT-737协同增强,并被泛半胱天冬酶抑制剂BOC-d-fmk或Bcl-XL的过表达抑制。进一步揭示,索拉非尼特异性地在BCR/ABL驱动的细胞中激活Bax和半胱天冬酶-3并降低线粒体膜电位。索拉非尼还抑制BCR/ABL诱导的其细胞底物的酪氨酸磷酸化及其在Ton.B210中的自身磷酸化。最终表明,在体外激酶测定中,索拉非尼抑制BCR/ABL及其E255K和T315I突变体的激酶活性。这些结果表明,索拉非尼至少部分地通过抑制BCR/ABL来激活线粒体介导的凋亡途径,从而诱导表达BCR/ABL的细胞凋亡。因此,索拉非尼可能为治疗Ph+白血病,特别是那些表达对伊马替尼和第二代BCR/ABL抑制剂完全耐药的T315I突变体的白血病,提供一种有效的治疗措施。