Application Of Proteomics Technology Screening And Analysis Gastric Cancer Cells Differentially Expressed Proteins

ObjectiveTwo-dimensional electrophoresis was used to investigate BGC-823cell, SGC-7901cell and GES-1cell (normal gastric cells). Screening and identifying the differentialexpressed proteins. Combined with MALDI-TOF-TOF-MS/MS Proteomics technology toidentify the identifying the differential expressed proteins. These proteins can be used astumor protein markers and can provide theoretical basis for the carcinogenic mechanism,early diagnosis and prognostic of gastric carcinoma.Methods1. BGC-823cell, SGC-7901cell and GES-1cell (normal gastric cells) were culturedin vitro, Extracted proteins respectively, used two-dimensional electrophoresis isolatedproteins, use PDQuest2-D Analysis Software8.0to screening differential proteins.2. Screening interested differences protein site to detect with MALDI-TOF-TOF-MS/MS Proteomics technology.Results1. We successfully separated proteins of BGC-823cell, SGC-7901cell and GES-1cell (normal gastric cells) by two-dimensional electrophoresis.1403±17proteins wereidentified in normal gastric cells,1407±15and1406±11proteins sites were identified inBGC-823cell and SGC-7901cell, respectively. From the results of paired BGC-823celland SGC-7901cell differences protein site found48differences protein site whichidentified22up-regulated proteins and26down-regulated proteins; From the results ofpaired GES-1cell and SGC-7901cell differences protein site found87differences proteinsite which identified47up-regulated proteins and40down-regulated proteins.2. In the above proteins, select ten protein sites respectively (fold change more thantwo times) and used MALDI-TOF-TOF-MS/MS proteomics technology to identified. Wesuccessfully identified16protein sites, including4up-regulated proteins and12 down-regulated proteins. These proteins can be roughly divided into: metabolic relevantenzymes, molecular partner, cytoskeletal protein process.Conclusions1. The result indicated that we successfully separate out and choose tumor proteinmarkers of related gastric cancer by two-dimensional electrophoresis. It can successfullyseparate1400protein sites in each gel figure.48differences protein site were found in theGES-1and BGC-823cells glue figure.87differences protein site were found in the GES-1and SGC7901cells gel figure.2.16proteins(fold change more than two times) were identified byMALDI-TOF-TOF-MS/MS proteomics technology, including4up-regulated proteins and12down-regulated proteins. it can divided into the following categories:(1) Metabolicrelevant enzymes: PPA1,PHGDH,LDHB,ENO1;(2) Molecular partner: HSP90β, HSP70;(3) Cytoskeletal proteins: CK7, Moesin, Lamin;(4) others: UBQLN1, ACTG1, GSTP1,PDIA3, WARS, CCT2and HNRNPF. These results lays foundation for the study of gastriccancer mechanism and tumor markers.3. In this stuty, HSP90β, UBQLN1and GSTP1appear protein modification, thespecific mechanism needs to be further study. There may be a close relationship betweenthe3proteins and gastric cancer development.