Rong Dongxiu, Lin Xiuxian, Luo Yanzhang, Mok Tin Seak, Wang Qing, Wang Haiyan, Zhang Tao
Department of Otorhinolaryngology, The First Affiliated Hospital, Jinan University, Guangzhou 510632, China.
Department of Otorhinolaryngology, Centro Hospitalar Conde de São Januário, Macao SAR 999078, China.
Transl Cancer Res. 2020 Jan;9(1):21-29. doi: 10.21037/tcr.2019.11.14.
We sought to determine the differences with respect to the proteome of nasopharyngeal tissues between patients with nasopharyngeal carcinoma (NPC) and healthy controls by using sequential windowed acquisition of all theoretical fragment ion mass spectra (SWATH-MS) and ingenuity pathway analysis (IPA). Our primary purpose was to identify specific protein markers that can be applied for diagnosis or treatment of NPC.
The CNE-1, CNE-2 and H1299 cell lines were cultured in stable isotope labeling of amino acids in cell culture (SILAC) medium for 10 generations to obtain labeled proteins. Thirty samples of NPC and 30 healthy control nasopharyngeal tissues were collected from the Department of Otolaryngology of the First Affiliated Hospital of Jinan University. Proteome of the nasopharyngeal tissues were analyzed and compared by SWATH-MS to identify differently expressed proteins. Further, extraction of target proteins and biological pathways was performed by IPA. Super-SILAC technique and liquid chromatography-tandem mass spectrometry were used to verify the reliability of the data obtained using SWATH-MS.
We identified 1,415 differentially expressed proteins between NPC patients and healthy controls. On IPA analysis, EIF2AK2 and MAPK1 proteins were found to be enriched in multiple biological pathways and functional networks.
The differentially expressed proteins EIF2AK2 and MAPK seem to play an important role in the biological network of NPC or may help discover the specific functional proteins of NPC. Further studies are required to identify the pathways and molecular mechanisms that underlie NPC.
我们试图通过使用所有理论碎片离子质谱的顺序窗口采集（SWATH-MS）和 Ingenuity 通路分析（IPA）来确定鼻咽癌（NPC）患者与健康对照者鼻咽组织蛋白质组的差异。我们的主要目的是识别可用于 NPC 诊断或治疗的特定蛋白质标志物。
将 CNE-1、CNE-2 和 H1299 细胞系在细胞培养中氨基酸的稳定同位素标记（SILAC）培养基中培养 10 代以获得标记蛋白。从暨南大学附属第一医院耳鼻喉科收集 30 份 NPC 样本和 30 份健康对照鼻咽组织。通过 SWATH-MS 分析和比较鼻咽组织的蛋白质组以鉴定差异表达的蛋白质。此外，通过 IPA 进行靶蛋白和生物通路的提取。使用超级 SILAC 技术和液相色谱 - 串联质谱来验证使用 SWATH-MS 获得的数据的可靠性。
我们鉴定出 NPC 患者和健康对照者之间有 1415 种差异表达的蛋白质。通过 IPA 分析，发现 EIF2AK2 和 MAPK1 蛋白在多个生物通路和功能网络中富集。
差异表达的蛋白质 EIF2AK2 和 MAPK 似乎在 NPC 的生物网络中起重要作用，或者可能有助于发现 NPC 的特定功能蛋白。需要进一步研究以确定 NPC 背后的通路和分子机制。
