定量磷酸化蛋白质组学分析鼻咽癌细胞中的 EGFR 信号通路。
Analysis of EGFR signaling pathway in nasopharyngeal carcinoma cells by quantitative phosphoproteomics.
机构信息
Key Laboratory of Cancer Proteomics of Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha 410008, China.
Key Laboratory of Allergy and Clinical Immunology, Department of Allergy, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou 510260, China.
出版信息
Proteome Sci. 2011 Jun 28;9:35. doi: 10.1186/1477-5956-9-35.
BACKGROUND
The epidermal growth factor receptor (EGFR) is usually overexpressed in nasopharyngeal carcinoma (NPC) and is associated with pathogenesis of NPC. However, the downstream signaling proteins of EGFR in NPC have not yet been completely understood at the system level. The aim of this study was identify novel downstream proteins of EGFR signaling pathway in NPC cells.
RESULTS
We analyzed EGFR-regulated phosphoproteome in NPC CNE2 cells using 2D-DIGE and mass spectrometry analysis after phosphoprotein enrichment. As a result, 33 nonredundant phosphoproteins including five known EGFR-regulated proteins and twenty-eight novel EGFR-regulated proteins in CNE2 were identified, three differential phosphoproteins were selectively validated, and two differential phosphoproteins (GSTP1 and GRB2) were showed interacted with phospho-EGFR. Bioinformatics analysis showed that 32 of 33 identified proteins contain phosphorylation modification sites, and 17 identified proteins are signaling proteins. GSTP1, one of the EGFR-regulated proteins, associated with chemoresistance was analyzed. The results showed that GSTP1 could contribute to paclitaxel resistance in EGF-stimulated CNE2 cells. Furthermore, an EGFR signaling network based on the identified EGFR-regulated phosphoproteins were constructed using Pathway Studio 5.0 software, which includes canonical and novel EGFR-regulated proteins and implicates the possible biological roles for those proteins.
CONCLUSION
The data not only can extend our knowledge of canonical EGFR signaling, but also will be useful to understand the molecular mechanisms of EGFR in NPC pathogenesis and search therapeutic targets for NPC.
背景
表皮生长因子受体（EGFR）在鼻咽癌（NPC）中通常过表达，与 NPC 的发病机制有关。然而，EGFR 在 NPC 中的下游信号蛋白在系统水平上尚未完全了解。本研究旨在鉴定 NPC 细胞中 EGFR 信号通路的新下游蛋白。
结果
我们使用 2D-DIGE 和质谱分析在磷酸蛋白富集后分析 NPC CNE2 细胞中的 EGFR 调节磷酸组。结果，鉴定出 33 个非冗余磷酸蛋白，包括 5 个已知的 EGFR 调节蛋白和 28 个新的 EGFR 调节蛋白，选择性验证了 3 个差异磷酸蛋白，并且显示 2 个差异磷酸蛋白（GSTP1 和 GRB2）与磷酸化 EGFR 相互作用。生物信息学分析表明，33 个鉴定蛋白中有 32 个含有磷酸化修饰位点，其中 17 个鉴定蛋白为信号蛋白。对 EGFR 调节蛋白之一 GSTP1 进行了分析，该蛋白与化疗耐药性有关。结果表明，GSTP1 可导致 EGF 刺激的 CNE2 细胞对紫杉醇产生耐药性。此外，使用 Pathway Studio 5.0 软件构建了基于鉴定的 EGFR 调节磷酸蛋白的 EGFR 信号网络，其中包括规范和新的 EGFR 调节蛋白，并暗示了这些蛋白的可能生物学作用。
结论
这些数据不仅可以扩展我们对规范的 EGFR 信号的认识，而且有助于理解 EGFR 在 NPC 发病机制中的分子机制，并寻找 NPC 的治疗靶点。
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