Mutation specific functions of EGFR result in a mutation-specific downstream pathway activation
Mutation specific functions of EGFR result in a mutation-specific downstream pathway activation
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DOI：
10.1016/j.ejca.2015.02.006
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发表时间：
2015-05-01
影响因子：
8.4
通讯作者：
French, Pim J.
中科院分区：
文献类型：
--
作者：
Erdem-Eraslan, Lale;Gao, Ya;French, Pim J.
背景：表皮生长因子受体（EGFR）在各种类型的癌症中经常发生突变。虽然所有的致癌突变都被认为是激活的，但不同的肿瘤类型有不同的突变谱。这可能是功能差异的基础上，这种肿瘤类型的特异性突变谱。（EGFR、EGFRvIII和EGFR-L 858 R）在结合配偶体、下游途径激活方面存在差异，（基因表达和磷蛋白），并对细胞生长和迁移的功能后果。使用生物素下拉和随后的质谱法，我们能够检测EGFR的突变特异性结合伴侣。使用邻近连接测定和/或蛋白质印迹法确认胞质分裂4（DOCK 4）、UDP-葡萄糖糖蛋白葡糖基转移酶1（UGGT 1）、MYC结合蛋白2（MYCBP 2）和Smoothelin（SMTN）的特异性结合。我们还证明，每个突变诱导一组特定的基因的表达，并且每个突变与特定的磷酸化模式。最后，我们证明使用稳定表达的细胞系，EGFRvIII和EGFL 858 R显示减少的生长和迁移相比，EGFR野生型expressing cells.Conclusion：我们的研究结果表明，有不同的EGFR突变之间的功能差异。不同突变之间的功能差异为开发突变特异性靶向治疗提供了依据。(C)2015爱思唯尔有限公司版权所有。
Background: Epidermal growth factor receptor (EGFR) is frequently mutated in various types of cancer. Although all oncogenic mutations are considered activating, different tumour types have different mutation spectra. It is possible that functional differences underlie this tumour-type specific mutation spectrum.Methods: We have determined whether specific mutations in EGFR (EGFR, EGFRvIII and EGFR-L858R) have differences in binding partners, differences in downstream pathway activation (gene expression and phosphoproteins), and have functional consequences on cellular growth and migration.Results: Using biotin pulldown and subsequent mass spectrometry we were able to detect mutation specific binding partners for EGFR. Differential binding was confirmed using a proximity ligation assay and/or Western Blot for the dedicator of cytokinesis 4 (DOCK4), UDP-glucose glycoprotein glucosyltransferase 1 (UGGT1), MYC binding protein 2 (MYCBP2) and Smoothelin (SMTN). We also demonstrate that each mutation induces the expression of a specific set of genes, and that each mutation is associated with specific phosphorylation patterns. Finally, we demonstrate using stably expressing cell lines that EGFRvIII and EGFL858R display reduced growth and migration compared to EGFR wildtype expressing cells.Conclusion: Our results indicate that there are distinct functional differences between different EGFR mutations. The functional differences between different mutations argue for the development of mutation specific targeted therapies. (C) 2015 Elsevier Ltd. All rights reserved.