Diindolylmethane Analogs Bind NR4A1 and Are NR4A1 Antagonists in Colon Cancer Cells
Diindolylmethane Analogs Bind NR4A1 and Are NR4A1 Antagonists in Colon Cancer Cells
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DOI：
10.1210/me.2014-1102
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发表时间：
2014-10-01
影响因子：
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通讯作者：
Safe, Stephen
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文献类型：
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作者：
Lee, Syng-Ook;Li, Xi;Safe, Stephen
1，1-二(3‘-吲哚)-1-(对取代苯基)甲烷(C-DIM)化合物在多种癌细胞系中显示出抗肿瘤活性，对羟基苯基类似物(DIM-C-pPhOH)在肺癌和胰腺癌细胞系中失活核受体4A1(NR4A1)。利用一系列14个不同的对取代苯基C-DIMS，我们证明了包括DIM-C-pPhOH在内的几个化合物直接与NR4A1的配体结合域相互作用。基于计算的分子模拟研究表明，DIM-C-pPhOH和相关化合物在NR4A1的配体结合口袋内具有高亲和力的相互作用，并且这些相同的化合物降低了结肠癌细胞中NR4A1依赖的反式激活，该构建含有3个串联的Nur77结合反应元件与荧光素酶报告基因相连。此外，我们还发现，通过RNA干扰(小干扰NR4A1)或用dim-C-pPhOH及其相关化合物处理NR4A1可抑制结肠癌细胞生长，诱导细胞凋亡，降低Survivin和其他Sp调节基因的表达，并抑制哺乳动物雷帕霉素信号转导的靶点。因此，C-DIMS，如DIM-C-pPhOH，直接与NR4A1结合，是结肠癌细胞中的NR4A1拮抗剂，它们的抗肿瘤活性部分是由于它们与核NR4A1的相互作用。
1,1-Bis(3'-indolyl)-1-(p-substituted phenyl) methane (C-DIM) compounds exhibit antineoplastic activity in multiple cancer cell lines and the p-hydroxyphenyl analog (DIM-C-pPhOH) inactivates nuclear receptor 4A1 (NR4A1) in lung and pancreatic cancer cell lines. Using a series of 14 different p-substituted phenyl C-DIMs, we show that several compounds including DIM-C-pPhOH directly interacted with the ligand binding domain of NR4A1. Computational-based molecular modeling studies showed high-affinity interactions of DIM-C-pPhOH and related compounds within the ligand binding pocket of NR4A1, and these same compounds decreased NR4A1-dependent transactivation in colon cancer cells transfected with a construct containing 3 tandem Nur77 binding response elements linked to a luciferase reporter gene. Moreover, we also show that knockdown of NR4A1 by RNA interference (small interfering NR4A1) or treatment with DIM-C-pPhOH and related compounds decreased colon cancer cell growth, induced apoptosis, decreased expression of survivin and other Sp-regulated genes, and inhibited mammalian target of rapamycin signaling. Thus, C-DIMs such as DIM-C-pPhOH directly bind NR4A1 and are NR4A1 antagonists in colon cancer cells, and their antineoplastic activity is due, in part, to their interactions with nuclear NR4A1.