Sirvent Audrey, Urbach Serge, Roche Serge
CNRS UMR5237, université de Montpellier 1 et 2, centre de recherche de biochimie macromoléculaire (CRBM), 34000 Montpellier, France.
CNRS UMR5203, Inserm U661, université de Montpellier 1 et 2, institut de génomique fonctionnelle (IGF), plate-forme de protéomique fonctionnelle, 34000 Montpellier, France.
Med Sci (Paris). 2014 May;30(5):558-66. doi: 10.1051/medsci/20143005020. Epub 2014 Jun 13.
Protein tyrosine kinases (TK) transmit intracellular signaling induced by many extracellular stimuli resulting in cell growth or adhesion. Deregulation of their activity leads to malignant cell transformation that plays an important role in human cancer. The signaling pathways involved in this oncogenic process are however only partially elucidated. Interestingly, SILAC-based quantitative proteomics allow the identification of the whole spectrum of TK substrates and the dynamic of phosphorylation state involved in oncogenic signaling. For example, this approach has highlighted the unsuspected complexity of the oncogenic signaling induced by the TK Src in colorectal cancer (CRC) cells. In this review, we describe a new SILAC-based technology applied to in vivo models of human tumors engrafted in nude mice. This method revealed significant differences between Src-oncogenic signaling of CRC cells in tumors and in culture. Finally, we discuss the interest of SILAC with recently described in vivo proteomic methods and in cancer, including the analysis of oncogenic signaling in tumor progression and the anti-tumoral activity of TK inhibitors in vivo.
蛋白质酪氨酸激酶（TK）传递由许多细胞外刺激诱导的细胞内信号，导致细胞生长或黏附。其活性失调会导致恶性细胞转化，这在人类癌症中起着重要作用。然而，参与这一致癌过程的信号通路仅得到部分阐明。有趣的是，基于稳定同位素标记氨基酸的细胞培养质谱定量蛋白质组学（SILAC）能够鉴定TK底物的全谱以及致癌信号中涉及的磷酸化状态动态变化。例如，这种方法突出了TK Src在结直肠癌（CRC）细胞中诱导的致癌信号的意外复杂性。在本综述中，我们描述了一种应用于裸鼠移植人肿瘤体内模型的基于SILAC的新技术。该方法揭示了肿瘤中CRC细胞与培养中的Src致癌信号之间的显著差异。最后，我们结合最近描述的体内蛋白质组学方法以及癌症研究中SILAC的应用价值进行讨论，包括肿瘤进展过程中致癌信号的分析以及TK抑制剂在体内的抗肿瘤活性。
