Disruption of Src function potentiates Chk1-inhibitor-induced apoptosis in human multiple myeloma cells in vitro and in vivo
Disruption of Src function potentiates Chk1-inhibitor-induced apoptosis in human multiple myeloma cells in vitro and in vivo
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DOI：
10.1182/blood-2010-06-291146
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发表时间：
2011-02-10
期刊：
影响因子：
20.3
通讯作者：
Grant, Steven
中科院分区：
文献类型：
--
作者：
Dai, Yun;Chen, Shuang;Grant, Steven
Ras/MEK/ERK通路激活代表了人多发性骨髓瘤（MM）细胞对检查点激酶1（Chk 1）抑制剂的重要代偿反应。为了研究Src在该事件中的功能作用和潜在的治疗意义，在体外和体内检查了Src和Chk 1抑制剂（例如，UCN-01或Chk 1 i）之间的相互作用。双Src/Abl抑制剂BMS 354825和SKI-606阻断Chk 1-受体诱导的细胞外信号调节激酶1/2（ERK 1/2）激活，显著增加MM细胞系和原发性CD 138（+）MM样本中与BimEL上调、p34（cdc 2）激活和DNA损伤相关的凋亡。Src功能缺失突变体（K297 R、K296 R/Y 528 F）或Src的shRNA敲低阻止了Chk 1抑制剂诱导的ERK 1/2激活并增加了细胞凋亡。相反，组成型活性Ras或丝裂原活化蛋白激酶/ERK激酶1（MEK 1）显着降低了Src抑制剂增强Chk 1抑制剂致死性的能力。此外，Src/Chk 1抑制剂共处理减弱了MM细胞产生血管内皮生长因子和其他血管生成因子（例如，ANG [血管生成素]、TIMP 1/2 [金属蛋白酶组织抑制剂1/2]和RANTES [受正常T细胞表达和分泌的活化调节]），并抑制体外血管生成。最后，在小鼠异种移植模型中，BMS 354825和UCN-01的联合给药抑制了人MM肿瘤生长，增加了细胞凋亡，并减少了血管生成。这些发现表明，Src激酶是Chk 1-激酶介导的Ras 3 ERK 1/2信号转导激活所必需的，并且该事件的破坏显著增强了Chk 1抑制剂在体外和体内的抗MM活性。（血。2011; 117（6）：1947-1957）
Ras/MEK/ERK pathway activation represents an important compensatory response of human multiple myeloma (MM) cells to checkpoint kinase 1 (Chk1) inhibitors. To investigate the functional roles of Src in this event and potential therapeutic significance, interactions between Src and Chk1 inhibitors (eg, UCN-01 or Chk1i) were examined in vitro and in vivo. The dual Src/Abl inhibitors BMS354825 and SKI-606 blocked Chk1-inhibitor-induced extracellular signal-regulated kinase 1/2 (ERK1/2) activation, markedly increasing apoptosis in association with BimEL up-regulation, p34(cdc2) activation, and DNA damage in MM cell lines and primary CD138(+) MM samples. Loss-of-function Src mutants (K297R, K296R/Y528F) or shRNA knock-down of Src prevented the ERK1/2 activation induced by Chk1 inhibitors and increased apoptosis. Conversely, constitutively active Ras or mitogen-activated protein kinase/ERK kinase 1 (MEK1) significantly diminished the ability of Src inhibitors to potentiate Chk1-inhibitor lethality. Moreover, Src/Chk1-inhibitor cotreatment attenuated MM-cell production of vascular endothelial growth factor and other angiogenic factors (eg, ANG [angiogenin], TIMP1/2 [tissue inhibitor of metalloproteinases 1/2], and RANTES [regulated on activation normal T-cell expressed and secreted]), and inhibited in vitro angiogenesis. Finally, coadministration of BMS354825 and UCN-01 suppressed human MM tumor growth in a murine xenograft model, increased apoptosis, and diminished angiogenesis. These findings suggest that Src kinase is required for Chk1-inhibitor-mediated Ras 3 ERK1/2 signaling activation, and that disruption of this event sharply potentiates the anti-MM activity of Chk1 inhibitors in vitro and in vivo. (Blood. 2011; 117(6): 1947-1957)