ARK5基因沉默联合5-氟尿嘧啶对胃癌细胞SGC7901生物学行为的影响.pdfVIP

Abstract ABSTRACT Objective: The tumor microenviroment is always in a ischemia, hypoxia and energy shortage condition due to the excessive proliferation of tumor cells.Meanwhile studies revealed that ARK5 was involved in the tolerance of tumor cells to local metabolic changes, and promoted tumor cells survival, invasion and metastasis. In the study, we will establish the lentiviral vectors of ARK5-RNAi to explore the effects of ARK5 gene silence combined with 5-fluorouracil on biological behaviors of gastric cancer cells. Methods: We designed three RNA interference (RNAi) sequences targeting the ARK5 gene and a negative control (NC) sequence and constructed the lentiviral vectors; The negative control lentivirus with GFP was used to infect the gastric cancer cells in order to get the optimum conditions of infection, and then we chosen the most effective ARK5-RNAi vector by Western Blot and transfected to SGC7901 cells; MTT colorimetric assay was used to test the cell growth inhibition rates in order to determine the concentration of 5-fluorouracil combined with ARK5 gene silence. The cultured SGC7901 gastric cancer cells were randomly divided into 6 groups: Control group, NC group, ARK5-RNAi group, Control+5-FU group, NC+5-FU group and ARK5-RNAi+5-FU group, we measured cell proliferation by MTT colorimetric assay, cell migration by scratch assay, cell invasion by transwell, cell apoptosis and cycle by flow cytometry, each group was repeated six times. Results: The result of gene sequencing showed that three ARK5-RNAi recombinant lentiviral vectors had been constructed successfully; When the SGC7901 cells were infected with viral particles in the presence of ENI.S. and Polybrene at a multiplicity of infection (MOI