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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.

Proteomics-Based Identification and Analysis of Proteins Associated with Helicobacter pylori in Gastric Cancer