【摘要】 背景:胃癌的发生是一个多因素多步骤的过程,幽门螺杆菌(H.pylori)感染在此过程中发挥重要作用。目的:建立胃癌和慢性胃炎H.pylori的双向凝胶电泳(2-DE)图谱,识别鉴定差异表达的蛋白质,探讨细菌本身因素在胃癌发病过程中的作用。方法:取H.pylori阳性的胃癌和慢性胃炎患者胃黏膜组织后分离培养H.pylori,收集菌体并采用BCA法行蛋白定量,以2-DE分离H.pylori蛋白,采用PDOuest软件分析差异表达的蛋白质点,应用电喷雾四极杆飞行时间串联质谱(O-TOF)进行鉴定,并在Mascot数据库中进行检索。结果:胃癌和慢性胃炎H.pylori总蛋白均获得分辨率高、重复性好的2-DE图谱。共6个蛋白质点在胃癌和慢性胃炎中存在表达差异,其中超氧化物歧化酶、尿素酶α亚单位、分子伴侣60 kDa在胃癌H.pylori菌株中高表达,而巯基过氧化物酶、二磷酸核苷激酶、S-核糖基同型半胱氨酸裂解酶在慢性胃炎H.pylori菌株中高表达。结论:成功建立了分辨率高且重复性好的胃癌和慢性胃炎H.pylori蛋白的2-DE图谱,并鉴定出两种疾病相关菌株中差异表达的蛋白质点,这些蛋白质点可能在胃癌的发生中起重要作用。更多还原

【Abstract】 Background:The pathogenesis of gastric cancer is a process with multiple factors and steps,and Helicobacter pylori(H.pylori) infection plays an important role in this process.Aims:To establish 2-dimensional gel electrophoresis (2-DE) maps of H.pylori in patients with gastric cancer and chronic gastritis,identify the differentially expressed proteins, and explore the role of bacterial factor in the pathogenesis of gastric cancer.Methods:H.pylori was isolated from biopsy specimen of gastric mucosa in patients with gastric cancer and chronic gastritis and then cultured.The whole-cell protein of H.pylori was extracted,and the protein concentration was measured by BCA method.Protein maps of H.pylori were established by 2-DE.Protein spots differentially expressed were analyzed by PDQuest software,and identified by electrospray ionization quadrupole time-of-flight mass spectrometry(Q-TOF).Then,they were retrieved by Mascot database. Results:High resolution and reproducible 2-DE maps of H.pylori were established in patients with gastric cancer and chronic gastritis.Six protein spots were differentially expressed between gastric cancer and chronic gastritis,3 protein spots of superoxide dismutase,urease subunit alpha and 60 kDa chaperonin were overexpressed in H.pylori of gastric cancer,and the other 3 protein spots of probable thiol peroxidase,nucleoside diphosphate kinase and S-ribosylhomocysteine lyase were overexpressed in H.pylori of chronic gastritis.Conclusions:High resolution and reproducible 2-DE maps of H.pylori have been established in patients with gastric cancer and chronic gastritis,and proteins spots differentially expressed between these two diseases are identified.The differential protein expression may play an important role in the pathogenesis of gastric cancer.更多还原