用比较蛋白质组学技术筛选口腔黏膜上皮细胞体外癌变的相关蛋白

Comparative proteomic analysis of differentially expressed proteins in the in vitro cellular carcinogenesis model of oral mucosal epithelial cells

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。

PURPOSE： To investigate the differential expressed proteins in the in vitro cellular carcinogenesis model of oral mucosal epithelial cells.METHODS： The in vitro cellular carcinogenesis model of oral mucosal epithelial cells was studied by comparative proteomic analysis.Two -dimensional gel electrophoresis was performed to separate the differentially expressed protein points with the PDQuest image analysis software.Mass spectrometry was then performed to identify the differentially expressed protein points.Gene Ontology Annotation was used to classify the known differentially expressed proteins.RESULTS： Using the two-dimensional gel electrophoresis and image analysis software of PDQuest,54 differentially expressed protein points were separated.Using the mass spectrometry of LC-MS/MS technique,45 different differentially expressed proteins were identified.According to the Gene Ontology Annotation,the known differentially expressed proteins were classified as cellular component,molecular function,and biological process.The most common types of cellular component were the cytoplasma and plasma membrane;the most common types of molecular function were phosphatase activity,catalytic activity,structural molecule activity,and calcium ion binding;the most common types of biological process were metabolic process,cell signaling and transduction,and cell adhesion and motility.CONCLUSIONS： The comparative proteomic analysis with two-dimensional gel electrophoresis and mass spectrometry could successfully separate and identify the differentially expressed proteins in the different cells of different stages of the in vitro cellular carcinogenesis model.