Drug resistance is dramatically restored by hedgehog inhibitors in CD34+ leukemic cells
Drug resistance is dramatically restored by hedgehog inhibitors in CD34+ leukemic cells
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DOI：
10.1111/j.1349-7006.2009.01111.x
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发表时间：
2009-05-01
期刊：
影响因子：
5.7
通讯作者：
Kato, Junji
中科院分区：
文献类型：
--
作者：
Kobune, Masayoshi;Takimoto, Rishu;Kato, Junji
在多种人类癌症和癌症干细胞中，已经描述了刺猬（Hh）信号的异常再激活。然而，Hh信号在白血病细胞调控中的作用仍不清楚。在这项研究中，我们评估了Hh通路激活对cd34（+）白血病细胞存活和耐药的影响。利用逆转录聚合酶链反应（RT-PCR）和Hh信号报告基因检测技术筛选白血病细胞系和原代白血病细胞中的Hh信号。我们发现Hh信号在几种人类急性髓性白血病（AML）细胞中是活跃的，特别是原代CD34（+）白血病细胞和细胞因子反应性CD34+细胞系，如Kasumi-1、Kasumi-3和TF-1。这些CD34+细胞表达下游效应物胶质瘤相关癌基因同源物（glii）1或GLI2，表明Hh信号传导活跃。此外，使用天然来源的Smoothened拮抗剂环巴胺、内源性Hh抑制剂刺猬相互作用蛋白或抗刺猬中和抗体抑制Hh信号在暴露48小时后诱导凋亡，尽管这些CD34（+）细胞系表现出对阿糖胞苷（Ara-C）的抗性。相比之下，环巴胺在缺乏Hh受体组分表达的U937和HL-60细胞系中没有影响生长或存活，证实了Hh抑制的作用是特异性的。此外，与10 μ M环巴胺联合可显著降低CD34（+）细胞系和原代CD34（+）白血病细胞对Ara-C的耐药性。这些结果表明Hh通路异常激活是一些CD34（+）髓系白血病细胞的一个特征，Hh抑制剂可能在AML的治疗中具有治疗作用。[j] .癌症科学2009；100:948-955。
Aberrant reactivation of hedgehog (Hh) signaling has been described in a wide variety of human cancers and in cancer stem cells. However, the contribution of Hh signaling to leukemic cell regulation has remained unclear. In this study, we assessed the possibility that Hh pathway activation contributes to the survival and drug resistance of cluster of differentiation (CD)34(+) leukemia cells. Hh signaling in leukemic cell lines and primary leukemic cells was screened by reverse transcription - polymerase chain reaction (RT-PCR) and a Hh signaling reporter assay. We found that Hh signaling is active in several human acute myeloid leukemia (AML) cells, especially primary CD34(+) leukemic cells and cytokine-responsive CD34+ cell lines such as Kasumi-1, Kasumi-3 and TF-1. These CD34+ cells express the downstream effectors glioma-associated oncogene homolog (GLI)1 or GLI2, indicative of active Hh signaling. Moreover, inhibition of Hh signaling with the naturally derived Smoothened antagonist cyclopamine, endogenous Hh inhibitor hedgehog-interacting protein or anti-hedgehog neutralizing antibody induced apoptosis after 48 h of exposure, although these CD34(+) cell lines exhibited resistance to cytarabine (Ara-C). In contrast, cyclopamine failed to affect growth or survival in U937 and HL-60 cell lines that lack expression of Hh receptor components, confirming that the effect of Hh inhibition is specific. Furthermore, combination with 10 mu M cyclopamine significantly reduced drug resistance of CD34(+) cell lines and primary CD34(+) leukemic cells to Ara-C. These results suggest that aberrant Hh pathway activation is a feature of some CD34(+) myeloid leukemic cells and Hh inhibitors may have a therapeutic role in the treatment of AML. (Cancer Sci 2009; 100: 948-955).