Aurora A, aurora B and survivin are novel targets of transcriptional regulation by histone deacetylase inhibitors in non-small cell lung cancer
Aurora A, aurora B and survivin are novel targets of transcriptional regulation by histone deacetylase inhibitors in non-small cell lung cancer
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Aurora A、Aurora B 和 survivin 是组蛋白脱乙酰酶抑制剂在非小细胞肺癌中转录调控的新靶点
DOI：
10.4161/cbt.7.9.6415
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发表时间：
2008-09-01
影响因子：
3.6
通讯作者：
Schrump, David S.
中科院分区：
文献类型：
--
作者：
Zhang, Xu-Hui;Rao, Mahadev;Schrump, David S.
背景：最近一项临床试验的活检分析表明，多肽FK228(DP)抑制肺癌细胞中Aurora激酶的表达。结果：肺癌细胞Aurora A和Aurora B的mRNA水平明显高于正常肺上皮细胞。DP、TSA和SAHA抑制Aurora A、Aurora B和Survivin表达的动力学在单个细胞系内非常相似，似乎与P53的表达状态一致。在用格尔达那霉素治疗后，没有观察到这些效果。抑制Aurora B转录的同时，H3K9Ac和H3K4me2激活标记减少，H3K9Me3以及MBD1、Mbd2和MBD3抑制标记在Aurora B最小启动子内积聚。敲除MBD1、-2或-3不能重复地阻断DP或TSA对Aurora或Survivin表达的抑制。DP和TSA降低Aurora激酶和Survivin的表达并改变其在肺癌细胞中的定位，导致肺癌细胞发生有丝分裂灾难。方法：应用定量RT-PCR技术检测肺癌细胞和正常呼吸道上皮细胞中Aurora A和Aurora B的表达水平。用这些方法以及免疫印迹法检测了DP、TSA、SAHA和格尔达霉素对肺癌细胞Auroras A/B及其相关基因/蛋白的表达。采用瞬时转染启动子-报告基因分析和染色质免疫沉淀(ChIP)技术检测DP对Aurora B启动子活性和染色质结构的影响。用共聚焦成像技术检测DP和TSA对肺癌细胞有丝分裂进程的影响。结论：涉及Aurora激酶和Survivin的新的转录调控机制可能参与了HDAC抑制剂对肺癌细胞的细胞毒作用。
Background: Analysis of biopsies from a recent clinical trial suggested that Depsipeptide FK228 (DP) inhibits Aurora kinase expression in lung cancer cells. The present study was undertaken to confirm and extend these observations.Results: Aurora A and B mRNA levels in lung cancer cells were considerably higher than levels in normal pulmonary epithelia. DP, TSA and SAHA inhibited Aurora A, Aurora B and survivin expression with kinetics that were remarkably similar within individual cell lines, and appeared to coincide with p53 expression status. These effects were not observed following treatment with geldanamycins. Inhibition of Aurora B transcription coincided with decreased H3K9Ac and H3K4Me2 activation marks, and accumulation of H3K9Me3, as well as MBD1, MBD2 and MBD3 repression marks within the minimal Aurora B promoter. Knockdown of MBD1, -2 or -3 did not reproducibly abrogate inhibition of Aurora or survivin expression by DP or TSA. DP and TSA decreased expression and altered localization of Aurora kinases and survivin, resulting in mitotic catastrophe in lung cancer cells.Methods: Aurora A, and Aurora B levels in lung cancer cells and normal respiratory epithelia were assessed using quantitative RT-PCR techniques. These methods, as well as as Western blots were used to examine expression of Auroras A/B, and several related genes/proteins in lung cancer cells exposed to DP, TSA, SAHA and geldanamycins. Transient transfection promoter-reporter assays, and chromatin immunoprecipitation (ChIP) techniques were used to examine DP-mediated changes in activity and chromatin structure of the Aurora B promoter. Confocal imaging techniques were used to examine the effects of DP and TSA on mitotic progression in lung cancer cells.Conclusions: Novel transcriptional regulatory mechanisms involving Aurora kinase and survivin appear to contribute to cytotoxicity mediated by HDAC inhibitors in lung cancer cells.