胃癌及慢性浅表性胃炎幽门螺杆菌临床株的分离及2-DE图谱的建立  

Establishment of 2-DE map of clinical strains of helicobacter pylori from gastric cancer and chronic superficial gastritis

作　　者：游运辉[1] 范学工[1] 刘萍[1] 刘洪波[1] 田雪飞[1] 

机构地区：[1]中南大学湘雅医院,湖南长沙410008

出　　处：《中国现代医学杂志》2009年第11期1680-1683,共4页China Journal of Modern Medicine

摘　　要：目的从临床标中分离HP,建立分别来源于胃癌及慢性浅表性胃炎Hp临床株的2-DE图谱,为分析鉴定差异蛋白质奠定基础。方法胃镜下钳取标本后分离、培养,并应用革兰染色及PCR鉴定所分离菌株。机械匀浆法将细菌破碎后加裂解液提取总蛋白,应用分步抽提法分离细菌蛋白,凝胶经银染显色后,用PDQuest图像分析软件进行凝胶图像分析。结果所分离菌株均为HP,获得背景清晰,分辨率高,重复性好的胃癌及慢性浅表性胃炎Hp临床株总蛋白2-DE图谱各3块,每张图谱约200个蛋白质点,大多数蛋白质点在位置和丰度上一致。结论成功建立了胃癌及慢性浅表性胃炎Hp临床株的2-DE图谱,并发现了14个差异表达的蛋白质点。[Objective] To isolate Helicobacter pylori strains from clinical specimens, establish 2-DE map of clinical strains of H. pylori separately from gastric cancer and chronic superficial gastritis, to lay the foundation for the analysis of differences in protein identification. [Methods] The specimens of Helicobacter pylori strains in human gastritis and gastric cancer were clamped under gastroscope and isolated and identified with Gram staining and PCR test. The bacteria were broken by Mechanical homogenate law, added lysate, and the total protein was extracted, via step-by-step extraction to separate bacterial protein, by silver staining gel color for later PDQuest gel image analysis software for image analysis. [Results] Helicobacter pylori strains were successfully isolated. Three 2-DE maps were of clinical strains from gastric cancer and chronic superficial gastritis H. pylori were obtained with background clear, high resolution, excellent repeatability. Each map are 200 protein spots, the majority of protein spots in the same location and breadth. [Conclusion] The 2-DE maps of clinical strains from gastric cancer and chronic superficial gastritis H. pylori were successfully established, and the 14 differentially expressed proteins were found.

关 键 词：胃癌 慢性浅表性胃炎 双向凝胶电泳 蛋白质组学 幽门螺杆菌 

分 类 号：R735.2[医药卫生—肿瘤]