Combining histone deacetylase inhibitor vorinostat with aurora kinase inhibitors enhances lymphoma cell killing with repression of c-Myc, hTERT, and microRNA levels.
Combining histone deacetylase inhibitor vorinostat with aurora kinase inhibitors enhances lymphoma cell killing with repression of c-Myc, hTERT, and microRNA levels.
复制标题
将组蛋白脱乙酰酶抑制剂伏立诺他与极光激酶抑制剂相结合，可通过抑制 c-Myc、hTERT 和 microRNA 水平来增强对淋巴瘤细胞的杀伤作用。
DOI：
10.1158/0008-5472.can-10-2259
复制
发表时间：
2011-06-01
期刊：
影响因子：
11.2
通讯作者：
Kirschbaum MH
中科院分区：
文献类型：
--
作者：
Kretzner L;Scuto A;Dino PM;Kowolik CM;Wu J;Ventura P;Jove R;Forman SJ;Yen Y;Kirschbaum MH
MK-0457和MK-5108是导致G2/M细胞周期停滞的新型极光激酶抑制剂（阿基）。使用MTS和膜联蛋白V测定法，单独使用药物或与伏立诺他（一种HDACi）联合使用，研究多种淋巴瘤细胞系的生长和存活，然后进行分子研究。100 - 500 nM的单独阿基导致约50%的细胞生长减少和10% - 40%的细胞凋亡。添加伏立诺他重新激活促凋亡基因并增强淋巴瘤细胞死亡。qPCR和免疫印迹显示，表观遗传和蛋白质乙酰化机制是这种活性的原因。联合药物治疗使促生存基因Bcl-XL和hTERT下调5倍，而促凋亡基因Bad和Bid上调3倍。p53肿瘤抑制因子通过响应伏立诺他的乙酰化增加和响应极光激酶A的Ser 315磷酸化减少而稳定。伏立诺他或阿伐他汀A降低Myc mRNA和蛋白以及Myc调节的microRNA。Myc是这些反应中的关键基因，因为Myc敲低与Myc拮抗剂Mxd 1的表达相结合，提高了细胞对任一阿基作用的敏感性。因此，HDACi伏立诺他导致转录和转录后变化以产生促凋亡环境，使细胞对有丝分裂特异性试剂如阿基敏感。
MK-0457 and MK-5108 are novel aurora kinase inhibitors (AKi) leading to G2/M cell cycle arrest. Growth and survival of multiple lymphoma cell lines were studied with either drug alone or in combination with vorinostat, an HDACi, using MTS and Annexin V assays, followed by molecular studies. Either AKi alone at 100 – 500 nM resulted in ~50% reduced cell growth and 10% – 40% apoptosis. Addition of vorinostat reactivated pro-apoptotic genes and enhanced lymphoma cell death. qPCR and immunoblotting revealed that epigenetic and protein acetylation mechanisms were responsible for this activity. The prosurvival genes Bcl-XL and hTERT were downregulated 5-fold by combination drug treatment, while the proapoptotic Bad and Bid genes were upregulated 3-fold. The p53 tumor suppressor was stabilized by an increased acetylation in response to vorinostat and a reduced Ser315 phosphorylation in response to aurora kinase A. Vorinostat or trichostatin A decreased Myc mRNA and protein as well as Myc-regulated microRNAs. Myc is a critical gene in these responses, as Myc knock-down combined with the expression of the Myc antagonist Mxd1, raised cell sensitivity to the effects of either AKi. Thus, the HDACi vorinostat leads to both transcriptional and post-transcriptional changes to create a pro-apoptotic milieu, sensitizing cells to mitosis-specific agents such as Aki’s.