Development of Biochemical Assays for the Identification of eIF4E-Specific Inhibitors
Development of Biochemical Assays for the Identification of eIF4E-Specific Inhibitors
复制标题
DOI：
10.1177/1087057112438554
复制
发表时间：
2012-06-01
影响因子：
--
通讯作者：
Magnaghi, Paola
中科院分区：
文献类型：
--
作者：
Visco, Carlo;Perrera, Claudia;Magnaghi, Paola
mRNA翻译的控制在细胞生长、增殖和分化中起关键作用，并且受到AKT和RAS致癌途径的严格调节。在此过程的调控中的关键参与者是mRNA 5'帽结合蛋白，真核翻译起始因子4 E（eIF 4 E）。eIF 4 E通过选择性地使有限的mRNA库能够翻译而导致恶性肿瘤，所述mRNA通常编码参与细胞周期进展、血管生成和转移的关键蛋白。一些数据表明，在肿瘤细胞系和异种移植模型中抑制eIF 4 E会损害肿瘤生长并诱导细胞凋亡;因此，eIF 4 E可以被认为是癌症治疗的有价值的靶标。靶向eIF 4 E的帽结合口袋应该代表抑制所有eIF 4 E细胞功能的方法。我们在这里提出的发展和验证不同的生化检测荧光偏振和表面等离子体共振技术的基础上。这些测定可以支持eIF 4 E抑制剂的高通量筛选、进一步改进和表征，以及针对真核细胞的另一种主要帽结合复合物CBP 80/CBP 20的选择性评估，总体上为eIF 4 E特异性抑制剂的开发提供了稳健的路线图。
Control of mRNA translation plays a critical role in cell growth, proliferation, and differentiation and is tightly regulated by AKT and RAS oncogenic pathways. A key player in the regulation of this process is the mRNA 5' cap-binding protein, eukaryotic translation initiation factor 4E (eIF4E). eIF4E contributes to malignancy by selectively enabling the translation of a limited pool of mRNAs that generally encode key proteins involved in cell cycle progression, angiogenesis, and metastasis. Several data indicate that the inhibition of eIF4E in tumor cell lines and xenograft models impairs tumor growth and induces apoptosis; eIF4E, therefore, can be considered a valuable target for cancer therapy. Targeting the cap-binding pocket of eIF4E should represent a way to inhibit all the eIF4E cellular functions. We present here the development and validation of different biochemical assays based on fluorescence polarization and surface plasmon resonance techniques. These assays could support high-throughput screening, further refinement, and characterization of eIF4E inhibitors, as well as selectivity assessment against CBP80/CBP20, the other major cap-binding complex of eukaryotic cells, overall providing a robust roadmap for development of eIF4E-specific inhibitors.