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目的:探讨口腔黏膜上皮细胞在体外癌变不同阶段表达的差异蛋白质。方法:以口腔黏膜上皮细胞体外癌变模型为对象,采用双向凝胶电泳技术和图像分析软件PDQuest分离和分析不同阶段细胞间的差异蛋白质点,采用LC-MS/MS质谱分析系统鉴定差异蛋白质点,采用Gene Ontology Annotation将已知差异蛋白质进行分类。结果:采用双向凝胶电泳技术和图像分析软件PDQuest,得到差异蛋白质点54个,采用LC-MS/MS质谱鉴定后,共得到候选差异蛋白质45个。根据Gene Ontology Annotation分类,按细胞组成分布最多的差异蛋白质位于细胞质和细胞膜,按分子功能分布最多的是磷酸酶活性、催化活性、结构分子功能和钙离子结合功能,按生物过程分布最多的是代谢过程、细胞信号传导和细胞黏附与运动。结论:比较蛋白质组学方法中的双向凝胶电泳和质谱技术,能够很好分离和鉴定口腔黏膜上皮细胞体外癌变模型中不同阶段细胞的差异蛋白质。
Objective: To investigate the differential proteins expressed in oral mucosal epithelial cells in different stages of carcinogenesis. METHODS: Two-dimensional gel electrophoresis (PDGE) and image analysis software PDQuest were used to separate and analyze the differential protein spots in different stages of cells. The differential protein spots were identified by LC-MS / MS mass spectrometry. Known differential proteins were categorized using Gene Ontology Annotation. Results: Two-dimensional gel electrophoresis (2D gel electrophoresis) and image analysis software PDQuest were used to obtain 54 differential protein spots. Forty-five protein spots were identified by LC-MS / MS. According to the Gene Ontology Annotation classification, the most disparate proteins distributed by cell composition are located in the cytoplasm and the cell membrane. The most widely distributed proteins are phosphatase activity, catalytic activity, structural molecular function and calcium ion binding function, and the most biologically distributed proteins are metabolic Process, Cell Signaling and Cell Adhesion and Exercise. CONCLUSIONS: By comparing two-dimensional gel electrophoresis and mass spectrometry techniques in proteomics, differential proteins in cells at different stages in the in vitro carcinogenesis model of oral mucosal epithelial cells can be well separated and identified.