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目的初步确定胃癌和消化性溃疡、胃炎幽门螺杆菌(Hp)的差异蛋白质。方法通过从胃镜活检胃黏膜组织中分离培养 Hp,应用裂解液、超声破碎法提取 Hp 菌体蛋白质后经 Bradford法进行菌体蛋白质定量,采用双向电泳方法获得 Hp 菌体蛋白质图像,运用 ImageMaster5.0版图像分析软件找到胃癌与消化性溃疡、胃炎 Hp 的差异蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)和四极杆飞行时间电喷雾串联质谱(Q-TOF)鉴定差异蛋白质。最后,通过WWW.matrixscience.com,从 mascot 数据库检索差异蛋白质。结果选择胃癌、消化性溃疡和胃炎 Hp各3例进行双向电泳,与胃溃疡和胃炎相比,胃癌 Hp 中有4个蛋白质点高表达,质潜鉴定结果为硫氧还蛋白、腺苷酸激酶、单链 DNA 结合蛋白和核糖体蛋白,50 S,L7/L12,其 Mowse 分值分别为94、286、139和132,序列覆盖率分别为77%、33%、33%和28%。结论硫氧还蛋白具有抗氧化和抑制细胞凋亡的功能,可能与 Hp 的致癌作用相关,蛋白质组学研究在 Hp 与胃癌关系的研究领域有广泛的应用前景。
Objective To determine the differentially expressed proteins of gastric cancer and peptic ulcer, Helicobacter pylori (Hp) in gastritis. Methods Hp was isolated and cultured from gastric mucosa by gastroscopic biopsy. Hp bacterial protein was extracted by ultrasonic disruption method, then the protein was quantified by Bradford method. Two-dimensional gel electrophoresis was used to obtain the protein image of H. pylori. Using ImageMaster5.0 The image analysis software was used to identify differential proteins between gastric cancer and peptic ulcer and gastritis Hp. The differential proteins were identified by using matrix-assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF) and quadrupole time of flight electrospray ionization tandem mass spectrometry (Q-TOF). Finally, differential proteins are retrieved from the mascot database at WWW.matrixscience.com. Results Three cases of gastric cancer, peptic ulcer and gastritis Hp were selected for two-dimensional electrophoresis. Compared with gastric ulcer and gastritis, four protein spots were highly expressed in gastric cancer Hp. The results of mass spectrometry were thioredoxin, adenylate kinase , Single-stranded DNA binding protein and ribosomal protein, 50 S, L7 / L12, with Mowse scores of 94, 286, 139 and 132, respectively. The sequence coverage was 77%, 33%, 33% and 28%, respectively. Conclusions Thioredoxin has the function of anti-oxidation and inhibition of apoptosis, which may be related to the carcinogenesis of Hp. Proteomics research has a wide range of application prospects in the field of Hp and gastric cancer.