摘要
目的:建立胃癌高、低发地区胃癌与慢性胃炎相关幽门螺杆菌(Helicobacter pylori,Hp)的双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)图谱,识别鉴定其差异表达蛋白质,探讨细菌本身因素在发病过程中的作用。方法:胃癌高发区(青海省西宁市)和胃癌低发区(北京市)的胃癌及慢性胃炎患者,胃镜下取胃黏膜组织,分离培养得到Hp临床株并冻存于-80℃。实验时将保存菌株复苏后,收集菌体并提取蛋白,利用双向凝胶电泳分离Hp蛋白,采用PDQuest软件分析、比较差异表达蛋白质点,利用四极杆飞行时间电喷雾串联质谱(four extremelystem flight time electrospray ionization tandem mass spectrometry,ESI-Q-TOF-MS)鉴定差异蛋白质点,并用Mascot数据库进行检索。结果:两地区胃癌及慢性胃炎相关Hp菌株总蛋白进行双向凝胶电泳,获得分辨率较高、重复性较好的2-DE图谱。鉴定出的9个差异蛋白质点中有4个在两地区胃癌菌株中均高表达,即尿素酶α、分子伴侣dnaK、DNA引导的RNA聚合酶α、超氧物岐化酶;2个在两地区慢性胃炎菌株中均高表达,即巯基过氧化物酶和核苷二磷酸激酶;此外还有3个蛋白质点仅在一个地区差异性表达,即分子伴侣60×103和无机焦磷酸酶仅在西宁市胃癌菌株中高表达,S-核糖基同型半胱氨酸裂解酶仅在北京市胃癌菌株中高表达。结论:胃癌高、低发区胃癌相关临床菌株与同一地区胃炎相关菌株蛋白质比较存在差异,但两地区2/3的差异蛋白质是一致的,胃癌高发区胃癌相关菌株高表达的差异蛋白质多于胃癌低发区,提示致胃癌的Hp菌株在不同地区其致胃癌的机制是相似的,但胃癌高发区胃癌相关菌株可能有强于低发区的致胃癌作用。
Objective：To identify the differentially expressed proteins of Helicobacter pylori（Hp） in patients with gastritis and gastric cancer from areas of high and low incidence of gastric cancer by 2-dimensional electrophoresis（2-DE）,and to discuss the role of bacterial factor in pathogenesis.Methods： Hp in the endoscopic biopsy specimens of gastric mucosa of patients with gastritis and gastric cancer from areas of high（Xining）and low（Beijing） incidence of gastric cancer,were separated,cultured and saved at-80 ℃.The bacteria were recovered.Then the whole-cell protein of the Hp were extracted and characterized by 2-DE.The different protein spots were analyzed by PDQuest analysis software and identified by electrospray ionization quadruple time-of-flight mass spectrometry（ESI-Q-TOF-MS）,and searched by the Mascot database.Results： Nine differentially expressed proteins were identified,and four protein spots were over expressed in the protein maps from gastric cancer in both areas,which were： Urease subunit alpha,chaperone protein dnaK,superoxide dismutase,DNA-directed RNA polymerase subunit alpha;two protein spots were over expressed in the protein maps from gastritis in both areas,which were： Probablethiol peroxidase,nucleoside diphosphate kinase;60×103 chaperonin,and inorganic pyrophosphatase were over expressed only in the protein map from gastric cancer in Xining;S-ribosyl homocysteinelyase was over expressed only in the protein map from gastric cancer in Beijing.Conclusion： There are differences between proteomic analyses of Hp in patients with gastritis and gastric cancer in areas of high and low incidents of gastric cancer,but 2/3 of the protein spots over expressed in the areas are consistent.The protein spots over expressed from gastric cancer in the area with high incidence of gastric cancer are more than in the area with low incidence of gastric cancer.For the Hp extracted from patients with gastric cancer,the mechanism of gastric cancer may be similar,but the role of the Hp from the area with high incidence of gastric cancer may be stronger.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2011年第6期827-832,共6页
Journal of Peking University:Health Sciences
基金
国家自然科学基金(30770980)资助~~
关键词
螺杆菌
幽门
蛋白质组
胃肿瘤
胃炎
Helicobacter pylori
Proteome
Stomach neoplasms
Gastritis
