| Objective: In this study, we applied the fluorescence differential in-gel electrophoresis (DIGE) technique to identify gastric cancer-specific protein markers, developing new potential therapeutic targets of the disease. Methods: Gastric cancer tissues and adjacent normal mucosa were examined by two-dimensional gel electrophoresis (2-DE) after labeled with CyDye DIGE Fluors Cy3, Cy5 and Cy2. Intensity changes of protein spots detected with statistical significance were identified by MALDI-TOF MS or MS/MS. Then selected proteins were analyzed by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Results: Intensity changes of 33 spots were detected with statistical significance. 9 protein spots of them up-regulated otherwise 24 down-regulated. And 22 of them were identified by MALDI-TOF MS or MS/MS successfully. The differentially expressed proteins could be divided into seven groups based on their functions: molecular chaperones, cystoskeleton proteins, metabolic enzymes, proteins associated with cell cycle, proteins associated with cell proliferation differentiation and apoptosis, and proteins associated with lipoid metabolism. Conclusions: Our results suggest that DIGE is a usfull technique for differential expressed proteins screening and analysis in gastric cancer tissures. MnSOD, mutant desmin, prostaglandin F synthase, HSP60, HSP27, eTEF 1 A 1 and so on differently expressed proteins may be usful tumor markers for gastric cancer. We found that there are weak relationship between protein and mRNA after the genes encoding differential proteins were analized transcriptionally.
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