Inhibition of mitotic kinase Aurora suppresses Akt-1 activation and induces apoptotic cell death in all-trans retinoid acid-resistant acute promyelocytic leukemia cells.
Inhibition of mitotic kinase Aurora suppresses Akt-1 activation and induces apoptotic cell death in all-trans retinoid acid-resistant acute promyelocytic leukemia cells.
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抑制有丝分裂激酶 Aurora 可抑制 Akt-1 激活并诱导全反式维甲酸耐药急性早幼粒细胞白血病细胞凋亡细胞死亡
DOI：
10.1186/1479-5876-9-74
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发表时间：
2011-05-21
影响因子：
7.4
通讯作者：
Liu Q
中科院分区：
文献类型：
--
作者：
Xu DR;Huang S;Long ZJ;Chen JJ;Zou ZZ;Li J;Lin DJ;Liu Q
Aurora激酶确保细胞周期中染色体的准确分离，维持细胞分裂中的遗传完整性。VX-680是一种小分子极光激酶抑制剂，可干扰有丝分裂进入和双极纺锤体的形成。在这里，我们评估VX-680作为一个潜在的代理人用于治疗全反式维甲酸（ATRA）耐药的急性早幼粒细胞白血病（APL）在体外。CD 11b表达用于通过流式细胞术评估细胞分化。免疫荧光染色分析细胞单极纺锤体的形成。MTT法检测细胞增殖;采用亚G1期细胞计数和Annexin V/PI双染法检测细胞凋亡。Hoechst 33342染色观察凋亡细胞核形态学改变。Western blot检测Aurora-A（Aur-A）的激活及参与细胞凋亡的信号通路。JC-1探针测定线粒体去极化。VX-680通过减少激活位点Thr 288的自磷酸化来抑制Aur-A，同时在对ATRA耐药的APL细胞系NB 4-R2中产生单极有丝分裂纺锤体。此外，我们发现VX-680抑制细胞增殖的MTT法评估。流式细胞术显示VX-680通过Sub-G1或Annexin V/PI分析以剂量和时间依赖性方式导致凋亡细胞死亡。Hoechst 33342染色代表VX-680处理的细胞中具有核碎裂的典型凋亡细胞。重要的是，VX-680对Aurora激酶的抑制抑制Akt-1活化并诱导线粒体去极化，这最终通过激活半胱天冬酶途径导致细胞凋亡，如通过增加NB 4-R2细胞中的半胱天冬酶原-3和聚ADP核糖聚合酶（PARP）的蛋白水解裂解所示。我们的研究提示有丝分裂Aurora激酶抑制剂在靶向ATRA耐药白血病细胞方面具有潜在的临床应用价值。
Aurora kinase ensures accurate chromosome segregation during cell cycle, maintaining genetic integrity in cell division. VX-680, a small-molecule Aurora kinase inhibitor, interferes with mitotic entry and formation of bipolar spindles. Here, we evaluated VX-680 as a potential agent for treatment of all-trans retinoid acid (ATRA)-resistant acute promyelocytic leukemia (APL) in vitro. CD11b expression was utilized to assess cell differentiation by flow cytometry. Immunofluorescence staining was conducted to analyze formation of cell monopolar spindle. Cell proliferation was evaluated by MTT assay. Sub-G1 population and Annexin V/PI staining were used to measure cell apoptosis. Hoechst 33342 staining was applied for identifying morphological changes in nucleus of apoptotic cell. Aurora-A (Aur-A) activation and the signaling pathways involved in apoptosis were detected by Western blot. JC-1 probe was employed to measure mitochondrial depolarization. VX-680 inhibited Aur-A by reducing autophosphorylation at the activation site, Thr288, accompanied by producing monopolar mitotic spindles in APL cell line NB4-R2 that was resistant to ATRA. In addition, we found that VX-680 inhibited cell proliferation as assessed by MTT assay. Flow cytometry showed that VX-680 led to apoptotic cell death in both dose- and time-dependent manners by either Sub-G1 or Annexin V/PI analysis. Hoechst 33342 staining represented typical apoptotic cells with nuclear fragmentation in VX-680 treated cells. Importantly, VX-680 inhibition of Aurora kinase suppressed Akt-1 activation and induced mitochondrial depolarization, which eventually resulted in apoptosis by activation of caspase pathway, as indicated by increasing proteolytic cleavage of procaspase-3 and poly ADP ribose polymerase (PARP) in NB4-R2 cells. Our study suggested potential clinical use of mitotic Aurora kinase inhibitor in targeting ATRA-resistant leukemic cells.
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影响因子：
11.4
作者：
Duprez, E;Benoit, G;Lanotte, M
通讯作者：
Lanotte, M
影响因子：
20.3
作者：
Giles, Francis J.;Cortes, Jorge;Freedman, Steven J.
通讯作者：
Freedman, Steven J.
影响因子：
11.2
作者：
Billottet, Clotilde;Banerjee, Lalita;Khwaja, Asim
通讯作者：
Khwaja, Asim
影响因子：
11.2
作者：
Briassouli, Paraskevi;Chan, Florence;Linardopoulos, Spiros
通讯作者：
Linardopoulos, Spiros
DOI：
10.1615/jenvironpatholtoxicoloncol.v28.i3.60
发表时间：
2009-01-01
影响因子：
2.4
作者：
Raghu, K. G.;Yadav, Govind Kumar;Bhadauria, Smrati
通讯作者：
Bhadauria, Smrati