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目的探讨幽门螺杆菌(Hp)感染与人胃腺癌上皮细胞(AGS)蛋白质组变化的关系。方法采用双向凝胶电泳分离Hp11638提取液与AGS细胞相互作用24 h后的细胞全蛋白样品,考马斯亮蓝染色,PD quest图像分析软件分析比较,识别差异蛋白,将差异蛋白进行胶内酶解,经基质辅助电离解析飞行时间质谱获得肽质量指纹图谱(PMF),通过搜索蛋白质数据库完成对差异蛋白质的鉴定。结果差异点切下酶解后经过基质辅助激光解析电离飞行时间质谱鉴定,共鉴定出7个蛋白;其中经细菌裂解液刺激后蛋白表达上调的有4种,分别是烯醇化酶1变异体、Septin家族成员1 CRA_a亚型、热休克蛋白gp96前体和阿片生长因子受体1;3种蛋白表达下调,分别是线粒体翻译单元延长因子EF-Tu前体、3-alpha羟类固醇脱氢酶Ⅱb型、1型蛋白磷酸酶调节亚基3D。结论幽门螺杆菌与AGS细胞相互作用的过程中,与肿瘤生物学行为相关的几种细胞蛋白表达上调;与调节细胞正常生长代谢相关的细胞蛋白表达下调。
Objective To investigate the relationship between Helicobacter pylori (Hp) infection and proteomic changes of human gastric adenocarcinoma epithelial cells (AGS). Methods Two-dimensional gel electrophoresis was used to separate the whole cell protein samples of Hp11638 solution and AGS cells for 24 hours. Coomassie brilliant blue staining and PD quest image analysis software were used to analyze and identify the differentially expressed proteins, Peptide mass fingerprinting (PMF) was obtained by matrix-assisted ionization time of flight mass spectrometry, and differential proteins were identified by searching the protein database. The results of different points after the enzyme digestion after matrix-assisted laser desorption ionization time of flight mass spectrometry identification, a total of seven proteins were identified; which were stimulated by bacterial lysate protein expression in four, namely enolase 1 variants, Septin family member 1 CRA_a subtype, heat shock protein gp96 precursor and opioid growth factor receptor 1; downregulation of three proteins, respectively, mitochondrial translation unit elongation factor EF-Tu precursor, 3-alpha hydroxysteroid dehydrogenase Ⅱ b Type, type 1 protein phosphatase regulatory subunit 3D. Conclusions During the interaction between Helicobacter pylori and AGS cells, the expression of several cellular proteins related to tumor biological behavior is up-regulated; the expression of cellular proteins related to normal growth and metabolism of cells is down-regulated.