Balch Curt, Naegeli Kaleb, Nam Seungyoon, Ballard Brett, Hyslop Alan, Melki Christina, Reilly Elizabeth, Hur Man-Wook, Nephew Kenneth P
Medical Sciences Program, Indiana University, Bloomington, IN, USA.
Cancer Biol Ther. 2012 Jun;13(8):681-93. doi: 10.4161/cbt.20086. Epub 2012 Jun 1.
Previously, we demonstrated potent antineoplastic activity of a distinctive histone deacetylase inhibitor (HDACI), AR42, against chemoresistant CP70 ovarian cancer cells in vitro and in vivo. Here, in follow-up to that work, we explored AR42 global mechanisms-of-action by examining drug-associated, genome-wide microRNA and mRNA expression profiles, which differed from those of the well-studied HDACI vorinostat. Expression of microRNA genes in negative correlation with their "target" coding gene (mRNA) transcripts, and transcription factor genes with expression positively correlated with coding genes having their cognate binding sites, were identified and subjected to gene ontology analyses. Those evaluations showed AR42 gene expression patterns to negatively correlate with Wnt signaling (> 18-fold induction of SFRP1), the epithelial-to-mesenchymal transition (40% decreased ATF1), and cell cycle progression (33-fold increased 14-3-3σ). By contrast, AR42 transcriptome alterations correlated positively with extrinsic ("death receptor") apoptosis (> 2.3-fold upregulated DAPK) and favorable ovarian cancer histopathology and prognosis. Inhibition of Wnt signaling was experimentally validated by: (1) > 2.6-fold reduced Wnt reporter activity; and (2) 36% reduction in nuclear, activated β-catenin. Likely AR42 induction of multiple (type I or type II autophagic) cell death cascades was further supported by 57% decreased reliance upon reactive oxygen, increased mitochondrial membrane disruption, and caspase independence, as compared with vorinostat. Taken together, we demonstrate distinct antineoplastic pathway alterations, in aggressive ovarian cancer cells, following treatment with a promising HDACI, AR42. These combined computational and experimental approaches may also represent a straightforward means for mechanistic studies of other promising antineoplastics, and/or the identification of agents that may complement epigenetic therapies.
此前,我们证明了一种独特的组蛋白去乙酰化酶抑制剂(HDACI)AR42在体外和体内对化疗耐药的CP70卵巢癌细胞具有强大的抗肿瘤活性。在此,作为该研究的后续,我们通过检测与药物相关的全基因组微小RNA和信使核糖核酸(mRNA)表达谱,探索了AR42的整体作用机制,这些表达谱与研究充分的HDACI伏立诺他不同。我们鉴定了与它们的“靶标”编码基因(mRNA)转录本呈负相关的微小RNA基因,以及与具有同源结合位点的编码基因表达呈正相关的转录因子基因,并进行了基因本体分析。这些评估表明,AR42基因表达模式与Wnt信号传导呈负相关(分泌型卷曲相关蛋白1诱导超过18倍)、上皮-间质转化(活化转录因子1减少40%)和细胞周期进程(14-3-3σ增加33倍)。相比之下,AR42转录组改变与外源性(“死亡受体”)凋亡呈正相关(死亡相关蛋白激酶上调超过2.3倍)以及与良好的卵巢癌组织病理学和预后相关。Wnt信号传导的抑制通过以下实验得到验证:(1)Wnt报告基因活性降低超过2.6倍;(2)核内活化β-连环蛋白减少36%。与伏立诺他相比,AR42可能诱导多种(I型或II型自噬)细胞死亡级联反应,这进一步得到以下证据支持:对活性氧的依赖性降低57%、线粒体膜破坏增加以及半胱天冬酶非依赖性。综上所述,我们证明了在侵袭性卵巢癌细胞中,用一种有前景的HDACI——AR42治疗后会出现明显的抗肿瘤途径改变。这些综合的计算和实验方法也可能代表了一种直接的手段,用于其他有前景的抗肿瘤药物的作用机制研究,和/或鉴定可能补充表观遗传疗法的药物。