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目的:筛选肺鳞癌淋巴结转移相关的蛋白质。方法:采用激光捕获显微切割技术(lasercapture microdissection,LCM)分别从肺鳞癌原发灶组织和匹配的淋巴结转移灶癌组织中切割纯化鳞癌细胞;应用二维凝胶电泳(two-dimensional electrophoresis,2-DE)分离LCM纯化细胞的蛋白质;图像分析识别差异表达的蛋白质点;基质辅助激光解吸电离飞行时间质谱鉴定差异蛋白质点,Western印迹验证差异蛋白Rho-GDIα在LCM纯化的肺鳞癌原发灶肿瘤细胞(lung primary tumor cells,LPTC)和匹配的淋巴结转移灶肿瘤细胞(lymph node metastatic tumor cells,LNMTC)中的表达。结果:建立了LCM纯化的LPTC和匹配的LNMTC的2-DE图谱,质谱鉴定了22个差异蛋白质,与LPTC相比,8个蛋白质在LNMTC中表达明显增高。结论:22个差异蛋白可能与肺鳞癌淋巴结转移有关,为筛选肺鳞癌转移标志物奠定了基础。
Objective: To screen the lymph node metastasis - associated proteins in lung squamous cell carcinoma. Methods: The squamous cell carcinoma cells were excised from primary squamous cell carcinoma tissue and matched lymph node metastatic carcinoma tissue by laser capture microdissection (LCM). Two-dimensional gel electrophoresis , 2-DE) were used to separate the proteins of LCM-purified cells; image analysis identified differentially expressed protein spots; matrix-assisted laser desorption / ionization time-of-flight mass spectrometry identified differential protein spots and Western blotting to verify the differential expression of Rho-GDIα in LCM- Expression of lung primary tumor cells (LPTCs) and matched lymph node metastatic tumor cells (LNMTCs). RESULTS: A 2-DE map of LCM-purified LPTC and matched LNMTC was established, and 22 differential proteins were identified by mass spectrometry. Eight proteins were significantly up-regulated in LNMTC compared with LPTC. Conclusion: The 22 differentially expressed proteins may be related to lymph node metastasis of squamous cell lung cancer, which lays the foundation for the screening of lung squamous cell carcinoma metastasis markers.