The Effect Of Interference Targeting TRAF1on The Apoptosis And Migration Of Gastric Cancer Cell

Background and objective:Gastric cancer is one of the most common malignancies in the world. A number of studies from home and abroad found that the occurrence of Gastric cancer has a close relationship with Helicobaeter pylori(H.pylori). Our group has found that if we foster Normal gastric epithelium cell GES-1with H.Pylor gathered from gastric cancer patient, the gene of TRAF1will be increased than before, and we found that the expression of gene TRAF1is upregulated in the tissue of gastric precancerous lesions and gastric cancer. It is found that the occurrence and development of a wide variety of tumor have relevant relations with TRAF1at present. Therefore, our subject intends to establish the model of gastric cancer cells with interference of TRAF1to learn the relation between the occurence and develop of gastric cancer, especially the gastric cancer correlacted with H.pylori. and TRAF1. Methods:At first, we constructed3pLVX-shRNA2-TRAF1-shRNAs expression vector targeting TRAF1according to the references. Then we detected the expression of TRAF1in BGC823, SGC7901, MGC803gastric cancer cell lines through the real-time PCR and Western-blot method, and to select the cell of highest expression of TRAF1to do later experiments. We transfered the3pLVX-shRNA-TRAF1-shRNAs into the cell selected before by using lipofectamine2000. Then we detected the expression levels of mRNA and protein before and after transfection by Real-time PCR and Western-blot, and select the strongest interference efficiency TRAF1shRNA to do the next experiment. When it is sure that TRAF1have been interfed and expressed successfully, we would test the cell proliferation activity through MTT assay and to analyze apoptosis through Flow cytometry assay, then test the cell migration using Transwell migration assay.Results:1. In BGC823, SGC7901, MGC803three gastric gastric cancer cell lines, the expression of TRAF1are all increased, and the highest expression of TRAF1is in BGC823gastric cell lines.2. In the3TRAF1shRNAs constructed accord to references, the strongest interference efficiency ShRNA is pLVX-shRNA2-TRAF1-shRNA2.3. When the gene TRAF1of BGC823has been interferenced, the cell’s growing power is weaken and the apoptosis rate is increased.4. When the TRAF1of BGC823has been interferenced, the cell migration has no significant differences.Conclusions:The expression of TRAF1was upregulated in BGC823,SGC7901, MGC803three gastric cancer cell lines, and the most obvious is in BGC823. Through interference targeting TRAF1we can find that TRAF1inhibit the apoptosis of BGC823cell. These researches may suggested that TRAF1is play a role of anti-apoptotic, and it may help to elucidate the molecular mechanism of gastric cancer...