Proteomics-Based Identification and Analysis of Proteins Associated with Helicobacter pylori in Gastric Cancer.
Proteomics-Based Identification and Analysis of Proteins Associated with Helicobacter pylori in Gastric Cancer.
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基于蛋白质组学的胃癌幽门螺杆菌相关蛋白质的鉴定和分析。
DOI：
10.1371/journal.pone.0146521
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Guan Z
中科院分区：
文献类型：
--
作者：
Zhou J;Wang W;Xie Y;Zhao Y;Chen X;Xu W;Wang Y;Guan Z
幽门螺杆菌（幽门螺杆菌）是一种螺旋形的革兰氏阴性细菌，可导致人类胃中最常见的慢性感染。诱导胃癌尚未完全了解。为了进一步表征幽门螺杆菌病毒，我们通过感染了胃癌细胞系SGC-7901和AGS，用CAGA+ H.幽门螺杆菌感染了三个细胞系，并用携带全长CAGA基因的载体转染SGC-7901。使用蛋白质组技术从三个细胞系中表达蛋白质，并通过LC-MS/MS选择了三种细胞系的10个差异蛋白通过Western印迹：β-肌动蛋白，L-肽脱氢酶（LDH），二氢丙酰胺脱氢酶（DLD），MRNA加工因子19同源因子（PRPF19），ATP合成酶，Calsodulin（CAM），P64 Clcp，Ran---菌，Ran-细菌，Ran-- GTPase激活蛋白（朗格），p43和钙网蛋白的检测。 Western印迹显示，与高基因表达相比，在胃癌组织和/或转移性淋巴结中，β-肌动蛋白，LDH，DLD，PRPF19和CAM基因的表达在胃癌组织和/或转移性淋巴结中下调。在胃癌组织中观察到的幽门螺杆菌感染。在幽门螺杆菌感染的幽门螺杆菌和表达CAGA的细胞中，在启动子-570和-170位置分别甲基化和-170位置。 。
Helicobacter pylori (H. pylori) is a spiral-shaped Gram-negative bacterium that causes the most common chronic infection in the human stomach. Approximately 1%-3% of infected individuals develop gastric cancer. However, the mechanisms by which H. pylori induces gastric cancer are not completely understood. The available evidence indicates a strong link between the virulence factor of H. pylori, cytotoxin-associated gene A (CagA), and gastric cancer. To further characterize H. pylori virulence, we established three cell lines by infecting the gastric cancer cell lines SGC-7901 and AGS with cagA+ H. pylori and transfecting SGC-7901 with a vector carrying the full-length cagA gene. We detected 135 differently expressed proteins from the three cell lines using proteome technology, and 10 differential proteins common to the three cell lines were selected and identified by LC-MS/MS as well as verified by western blot: β-actin, L-lactate dehydrogenase (LDH), dihydrolipoamide dehydrogenase (DLD), pre-mRNA-processing factor 19 homolog (PRPF19), ATP synthase, calmodulin (CaM), p64 CLCP, Ran-specific GTPase-activating protein (RanGAP), P43 and calreticulin. Detection of the expression of these proteins and genes encoding these proteins in human gastric cancer tissues by real-time PCR (RT-qPCR) and western blot revealed that the expression of β-ACTIN, LDH, DLD, PRPF19 and CaM genes were up-regulated and RanGAP was down-regulated in gastric cancer tissues and/or metastatic lymph nodes compared to peri-cancerous tissues. High gene expression was observed for H. pylori infection in gastric cancer tissues. Furthermore, the LDH, DLD and CaM genes were demethylated at the promoter -2325, -1885 and -276 sites, respectively, and the RanGAP gene was highly methylated at the promoter -570 and -170 sites in H. pylori-infected and cagA-overexpressing cells. These results provide new insights into the molecular pathogenesis and treatment targets for gastric cancer with H. pylori infection.