人胃黏膜上皮细胞与胃癌细胞间酪氨酸磷酸化的比较蛋白质组学研究

Proteomics Analysis of Tyrosine Phosphorylation Between Gastric Mucosa Epithelial Cells and Gastric Cancer Cells

比较人正常胃黏膜上皮细胞GES-1与人胃癌细胞SGC-7901间酪氨酸磷酸化蛋白质的差异,筛选差异磷酸化蛋白质分子,为揭示胃癌发生发展的分子机制提供新的理论依据。采用免疫沉淀方法从人胃黏膜上皮细胞GES-1与人胃癌细胞SGC-7901总蛋白质中免疫沉淀出酪氨酸磷酸化蛋白质,用SDS-PAGE和二维凝胶电泳技术分离沉淀出的酪氨酸磷酸化蛋白质,银染,差异蛋白点进行胶内酶解,采用MALDI-TOF/TOF-MS质谱进行差异蛋白质鉴定。结果显示获得了7个差异酪氨酸磷酸化蛋白质,这些蛋白质涉及细胞骨架、细胞调控等。通过比较正常胃黏膜上皮细胞与胃癌细胞内酪氨酸磷酸化蛋白质的差异,筛选获得7个差异酪氨酸磷酸化蛋白质分子,有助于深入研究胃癌发生发展的分子机制,进而为胃癌的早期诊断和防治提供新的理论依据和作用靶标。

It was to compare differenti tyrosine-phosphoproteome between gastric mucosa epithelial GES-1 cells and gastric cancer SGC-7901 ceils, and elucidate the molecular mechanisms of gastric carcinogenesis. Total proteins were extracted from gastric mucosaepithelial GES-1 cells and gastric cancer SGC-7901 cells. Tyrosine-phosphorylated proteins in gastric mueosaepithelial GES-1 cells and gastric cancer SGC-7901 cells were immunoprecipitated by using immobilized phospho-tyrosine antibody, respectively. The immunoprecipitated tyrosine-phophorylated proteins were separated by SDS-PAGE or two dimensional gel electrophoresis （2-DE）. The differential protein spots were in-gel digested with trypsin, and identified with MALDI-TOF/TOF-MS. Results indicated that 7 different tyrosinephosphorylated proteins which were involved in cytoskeletons, regulation of cells and other aspects,were identified between gastric mucosa epithelial GES-1 cells and human gastric cancer SGC-7901 cells. It would be helpful to further define gastric carcinogenesis and provide the novel target proteins for early diagnosis and treatment of gastric cancer.