Screening Gastric Cancer Metastasis-associated Protein And Gene By Nude Mice Animal Models

ObjectiveObtain the pre and post metastatic gastric carcinoma by establishing a nude micemodel bearing a transplantable gastric carcinoma. Using two-dimensional electrophoresisto separate and screen differential proteins of the pre and post metastatic gastric carcinoma.Difference expression gene was screened by gene chip. It can provide theoretical basis forthe mechanism of gastric cancer metastasis and which is reducing the mortality of gastriccancer by targeted therapy.Methods1. BGC-823was cultured in vitro. Collecting cells and injecting them into nude micesubcutaneously. After tumor formation, planting small solid tumor that is passed severaltimes into stomach wall of nude mice and waiting them to be transferred.2. Obtain the pre and post metastatic gastric carcinoma(including tissue in gastricwall before and after metastasis, metastatic tissue in liver and mesenteric lymph nodes).Protein is extracted, and then separated by2-D electrophoresis. Using PDQest8.0softwareto analyse image and searching for different protein. Using4800plus MALDI TOF/TOFmass spectrometer to identify protein after synthesizing three group of experimentalresults.3. The gene expression profiling of tumor tissue before and after the transfer of thenude stomach was screened. The monochrome Cy3labeling and microarray hybridizationafter labeling, the fluorescence signal obtained different genes, then the fluorescence signalinto a digital signal. After statistical analysis of differentially expressed genes.Results1. Nude mice model with transplantable gastric carcinoma is successfully established.Metastatic gastric carcinoma was found in lobes of liver and mesenteric lymph nodes afterbeen planted9-10weeks in some mice.2. Different proteins were separated by using2-D electrophoresis from gastric tissue in gastric wall before and after metastasis, metastatic tissue in liver and mesenteric lymphnodes. Then we found82different protein sites between the pre metastaticgastric carcinoma and the post metastatic gastric carcinoma, including40up-regulatedproteins and42down-regulated proteins. There are120different protein sites between thepre metastatic gastric carcinoma and metastatic tissue in mesenteric lymph nodes,including76up-regulated proteins and44down-regulated proteins. There are63differentprotein sites between the post metastatic gastric carcinoma and metastatic tissue inmesenteric lymph nodes, including41up-regulated proteins and22down-regulatedproteins.Using4800plus MALDI TOF/TOF mass spectrometer, we triumphantly identifyingmany proteins, they are HSP9STRESS-70; Stmn1,2; Ezrin; Moesin; Anxa3; Calm1,2,3;Prdx4; Ranbp1; Npm1; Clicl; Actg1,g2, β; Hnrnph1; Capg; Apoal; Trf; PPa1; Enol; Aldoa;Pgk1; Try5.3.By gene chip to screen out at45,000genes, we found that there are2134geneexpression changes, including1078up-regulated genes and1056down-regulated genes.There are17metastasis related gene,7genes were up-regulated,10genes were downregulated.ConclusionSuccessfully prepared animal model of gastric cancer metastasis in nude mice, andmetastasis of gastric cancer screening of proteins and genes laid the foundation for furtherstudy of the mechanism of gastric cancer metastasis.