The Effect Of Interference Targeting TRAF1on The Apoptosis And Migration Of GES-1with Hp Infection

Background and objective:Helicobacter pylori is a class I carcinogens. Our group has found that if we foster Normal gastric epithelium cell GES-1with H.Pylori gathered from gastric cancer patient, the gene of TRAF1will be increased than before, and we found that if we interfere the expression of gene TRAF1in gastric cancer cell line, the growth activity of cell will be wesken, and the apoptoses will be increased significantly. Therefore, our subject intends to establish the model of GES-1infected by H.pylori with interference of TRAF1to learn the relation between the occruence and develop of gastric cancer correlacted with H.pylori.Methods:On the basis of preliminary experiments, GES-1cells were fostered with the H.pylori strains (2strains), the strains has the most significant proliferation effect to cells, after24hours, we will checked the expression of gene TRAF1by real-time PCR and Wsetern-blot method. Then we transferred the pLVX-shRNA2-TRAF1-shRNA2into the cell by using lipofectamine2000, and detected the expression levels of mRNA and protein of gene TRAF1after transfection by real-time PCR and Western-blot. When it is sure that TRAF1have been interfed and expressed successfully, we would test the cell proliferation actieity through MTT assay and to analyze apoptosis through Flow cytometry assay, then test the cell migration using Transwell migration assay and the scratch test.Results:1. Cocultivation2H.pylori strains with GES-1cells, the expression of TRAF1is increased; and pLVX-shRNA2-TRAF1-shRNA2has an obvious interference efficiency on gene TRAF1in co-culture cells. 2. When the gene TRAF1of GES-1infected with H.pylori has been interferenced, the cell’s growing power is weaken and the apoptosis rate is increased.3. When the gene TRAF1of GES-1infected with H.pylori has been interferenced, the cell’s migration has no significant differences.Conclusions:The expression of TRAF1was upregulated in GES-1cells cocultivated with H.pylori. Through interference targeting TRAF1we can find that TRAF1inhibit the apoptosis of co-culture cells. These researches may suggested that TRAF1play a role of anti-apoptotic in gastric cancer correlacted with H.pylori, and it may help to elucidate the molecular mechanism of gastric cancer correlacted with H.pylori.