幽门螺杆菌与胃上皮细胞不同作用时间点蛋白质组的差异分析  

Sequential proteomic characteristics of AGS cells co-cultured with H pylori strain

作　　者：宋衍燕[1] 赵飞[1] 肖迪[1] 马广源[1] 孟凡亮[1] 何丽华[1] 张建中[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所,北京市102206

出　　处：《世界华人消化杂志》2008年第12期1260-1265,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.39870032;"十一五"国家科技支撑计划重点资助项目;No.2007BAI04B02~~

摘　　要：目的:分析幽门螺杆菌(Hpylori)与人胃腺癌上皮细胞(humangastric adenocarcinoma epithelial,AGS)的相互作用蛋白质组学差异.方法:采用双向凝胶电泳分离Hpylori26695株与AGS细胞相互作用0.5、2、4h时间点的细胞和黏附菌的全蛋白样品,考马斯亮兰染色,ImageMster2D图像分析软件比较分析,识别差异蛋白,将差异蛋白点进行胶内酶解,经基质辅助电离解析飞行时间质谱(MALDI-TOF-TOF/MS)获得肽质量指纹图谱(PMF),通过搜寻蛋白质数据库完成对差异蛋白的鉴定.结果:对0.5、2、4h时间点图谱差异蛋白点分析共发现主要差异蛋白点66个,对其中34个(共对应16种蛋白)最终完成准确鉴定,2h后8种蛋白开始上调,其中2种是细胞来源,6种Hpylori来源;4h时4种细胞来源蛋白下调,并有4种新的蛋白开始被观察到.在共感染的早期,细胞的琥珀酸脱氢酶、Hpylori的抗氧化酶类、黏附素以及HSP60表达上调.感染晚期主要是细胞来源的亲环素A、新生多肽相关复合物α多肽开始表达,Hpylori来源的尿素酶、非血红素铁蛋白等蛋白明显出现在黏附菌成分中.结论:Hpylori与AGS细胞相互作用过程中,细菌和细胞蛋白表达均存在明显时序性变化,早期相互作用则主要表现为以与黏附等有关的蛋白表达变化,后期向有利细菌存活和增殖的方向发展,并呈现出与免疫逃逸和病理损伤相关的变化.AIM： To analyze the differential expression of proteome in human gastric adenocarcinoma epithelial AGS cells co-cultured with H pylori. METHODS： The samples of AGS cells co- cultured with H pylori 26695 strain at three time-points （0.5, 2 and 4 h） were collected and separated by 2-dimensional polyacrylamide gel electrophoresis （2-DE） technique and computerassisted image analysis was used to analyze the differential proteomic expression. The significantly differentially expressed proteins were recognized and identified with the 4700 proteomics discovery system. RESULTS： There were 66 protein spots that were significantly differentially expressed at different time-points, 34 （corresponding to 16 kinds of proteins） of which were identified with matrix-assisted laser desorption/ionization time- of-flight/time-of-flight （MALDI-TOF-TOF）. At 2-h time-point, 8 kinds of proteins were up-regulated obviously, of which 2 originated from the cells and 6 from H pylori. At the 4-h time-point, 4 kinds of cell-originated proteins were downregulated, and 4 kinds of new proteins were observed. Succinate dehydrogenase iron-sulfur subunit, HpaA, HSP 60 and peroxiredoxin were up-regulated at the early stage of co-infection. However, at the late stage of co-infection, two cell-originated proteins （cyclophilin A, nascentpolypeptide-associated complex alpha polypeptide） and two H pylori-originated proteins （urease, non-heme iron protein） were found to express CONCLUSION： During the interaction between AGS cells and H pylori, the expression of proteins is associated with adhesion change in the early stage, followed by a favorable alteration in the survival and proliferation of H pylori as well as immunologic escape and pathological erosion.

关 键 词：幽门螺杆菌 人胃腺癌上皮细胞 共培养 蛋白质组 

分 类 号：R730.43[医药卫生—肿瘤] R378.99[医药卫生—临床医学]