RNAi-mediated silencing of p190Bcr-Abl inactivates Stat5 and cooperates with imatinib mesylate and 17-allylamino-17-demetoxygeldanamycin in selective killing of p190Bcr-Abl-expressing leukemia cells
RNAi-mediated silencing of p190Bcr-Abl inactivates Stat5 and cooperates with imatinib mesylate and 17-allylamino-17-demetoxygeldanamycin in selective killing of p190Bcr-Abl-expressing leukemia cells
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DOI：
10.1038/leu.2008.60
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发表时间：
2008-06-01
期刊：
影响因子：
11.4
通讯作者：
Tojo, A.
中科院分区：
文献类型：
--
作者：
Futami, M.;Hatano, T.;Tojo, A.
190 kD（p190）和210 kD（p210）Bcr-Abl蛋白参与了Ph（+）白血病的病理生理过程。我们应用RNA干扰（RNAi）技术特异性杀伤p190（+）细胞，并分别确定了p190的BCR区、连接区和ABL区基因沉默的最佳序列。然后，用编码这些shRNA的慢病毒载体感染p190（+）和p210（+）细胞，导致p190（+）细胞的有效杀伤，而p210（+）细胞仅对shBCR和shABL敏感。在p190转化的Ba/F3细胞中，p190的沉默特异性抑制死亡前Stat 5的酪氨酸磷酸化，但不影响Jak 2、Akt或MEK 1/2的磷酸化。相反，通过用17-烯丙基氨基-17-去甲氧基格尔德霉素（17-AAG）处理下调p190与Jak 2、Akt和MEK 1/2的蛋白水平降低相关。靶向p190的shRNA与伊马替尼和17-AAG在Ba/F3-p190 wt和伊马替尼耐药Ba/F3 p190 Y253 H细胞的生长抑制中相加地协同作用。总的来说，RNAi介导的p190沉默是一个有前途的选择，既描绘信号转导和治疗应用于190（+）白血病。
The 190 kD (p190) and 210 kD (p210) Bcr-Abl proteins are responsible for the pathophysiology of Philadelphia chromosome (Ph)(+) leukemia. We applied RNA interference (RNAi) to specific killing of p190(+) cells, and determined the optimal sequences for gene silencing in the BCR, junctional and ABL regions of p190, respectively. Then, p190(+) and p210(+) cells were infected with lentiviral vectors encoding these shRNAs, resulting in efficient killing of p190(+) cells, while p210(+) cells were only sensitive to shBCR and shABL. In p190-transformed Ba/F3 cells, silencing of p190 specifically inhibited tyrosine phospohorylation of Stat5 prior to their death, but did not affect phosphorylation of Jak2, Akt or MEK1/2. In contrast, downregulation of p190 by their treatment with 17-allylamino-17demetoxygeldanamycin (17-AAG) was associated with reduced protein levels of Jak2, Akt and MEK1/2. shRNA targeting p190 collaborated additively with imatinib and 17-AAG in growth inhibition of Ba/F3-p190wt and imatinib-resistant Ba/F3p190Y253H cells. Collectively, RNAi-mediated silencing of p190 is a promising option both for delineating signal transduction and for therapeutic application in 190(+) leukemia.