Department of Hematology and Oncology, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871, USA.
J Biol Chem. 2010 Oct 8;285(41):31774-82. doi: 10.1074/jbc.M110.118653. Epub 2010 Jul 27.
BCR-ABL is a causative tyrosine kinase (TK) of chronic myelogenous leukemia (CML). In CML patients, although myeloid cells are remarkably proliferating, erythroid cells are rather decreased and anemia is commonly observed. This phenotype is quite different from that observed in polycythemia vera (PV) caused by JAK2 V617F, whereas both oncogenic TKs activate common downstream molecules at the level of hematopoietic stem cells (HSCs). To clarify this mechanism, we investigated the effects of BCR-ABL and JAK2 V617F on erythropoiesis. Enforced expression of BCR-ABL but not of JAK2 V617F in murine LSK (Lineage(-)Sca-1(hi)CD117(hi)) cells inhibited the development of erythroid cells. Among several signaling molecules downstream of BCR-ABL, an active mutant of N-Ras (N-RasE12) but not of STAT5 or phosphatidylinositol 3-kinase (PI3-K) inhibited erythropoiesis, while N-RasE12 enhanced the development of myeloid cells. BCR-ABL activated Ras signal more intensely than JAK2 V617F, and inhibition of Ras by manumycin A, a farnesyltransferase inhibitor, ameliorated erythroid colony formation of CML cells. As for the mechanisms of Ras-induced suppression of erythropoiesis, we found that GATA-1, an erythroid-specific transcription factor, blocked Ras-mediated mitogenic signaling at the level of MEK through the direct interaction. Furthermore, enforced expression of N-RasE12 in LSK cells derived from p53-, p16(INK4a)/p19(ARF)-, and p21(CIP1/WAF1)-null/wild-type mice revealed that suppressed erythroid cell growth by N-RasE12 was restored only by p21(CIP1/WAF1) deficiency, indicating that a cyclin-dependent kinase (CDK) inhibitor, p21(CIP1/WAF1), plays crucial roles in Ras-induced suppression of erythropoiesis. These data would, at least partly, explain why respective oncogenic TKs cause different disease phenotypes.
BCR-ABL 是慢性髓系白血病(CML)的致病酪氨酸激酶(TK)。在 CML 患者中,尽管髓系细胞明显增殖,但红细胞减少,贫血很常见。这种表型与 JAK2 V617F 引起的真性红细胞增多症(PV)完全不同,而这两种致癌 TK 在造血干细胞(HSCs)水平上激活共同的下游分子。为了阐明这一机制,我们研究了 BCR-ABL 和 JAK2 V617F 对红细胞生成的影响。在小鼠 LSK(谱系(-)Sca-1(hi)CD117(hi))细胞中强制表达 BCR-ABL 但不表达 JAK2 V617F 会抑制红细胞的发育。在 BCR-ABL 下游的几种信号分子中,一种活性突变的 N-Ras(N-RasE12)而非 STAT5 或磷脂酰肌醇 3-激酶(PI3-K)抑制红细胞生成,而 N-RasE12 增强了髓系细胞的发育。BCR-ABL 激活 Ras 信号的强度强于 JAK2 V617F,法呢基转移酶抑制剂曼马霉素 A 抑制 Ras 可改善 CML 细胞的红细胞集落形成。至于 Ras 诱导的红细胞生成抑制的机制,我们发现,红细胞特异性转录因子 GATA-1 通过直接相互作用在 MEK 水平上阻断 Ras 介导的有丝分裂信号。此外,在源自 p53-、p16(INK4a)/p19(ARF)-和 p21(CIP1/WAF1)-缺失/野生型小鼠的 LSK 细胞中强制表达 N-RasE12 表明,N-RasE12 抑制红细胞生长的作用仅通过 p21(CIP1/WAF1)缺陷得到恢复,表明细胞周期蛋白依赖性激酶(CDK)抑制剂 p21(CIP1/WAF1)在 Ras 诱导的红细胞生成抑制中发挥关键作用。这些数据至少部分解释了为什么各自的致癌 TK 导致不同的疾病表型。