Effects Of BRMS1 Gene Transfection On The Vitro Invasion Ability And Proliferation Of SGC7901 Gastric Cancer Cells

Purpose : Will eukaryotic expression vector pcDNA3.1 (-) B / myc-BRMS1 transfected into SGC7901 gastric cancer cells, BRMS1 observed in vitro gene SGC7901 cell proliferation and invasion capacity of.Method:Conventional culture of gastric cancer cells SGC7901, RT-PCR method to detect SGC7901 gastric cancer cells BRMS1 mRNA expression; With liposomes LIPOFECTAMINE 2000 will pcDNA3.1/myc-his (-) B and pcDNA3.1/myc-BRMS1 transfected into gastric cancer cells SGC7901, 48 hours after transfection with 300 ug / ml of the G418 pressure screening 28 days, Resistance to be formed after the cloning of human beings with 96-cell culture plate limited dilution pick from monoclonal, Monoclonal will use 170 ug / ml of the G418 to maintain and expand screening training; After the train was amplified by RT-PCR and Western Blot methods compare before and after the transfer BRMS1 changes in gene expression, From transfected BRMS1 the most obvious changes in gene expression in the cell cloning method drawn by MTT cell growth curve, Boyden chamber with improved lines of cells in vitro invasion and metastasis experimentResults : SGC7901 gastric cancer cells in gene expression without BRMS1; Transfection eukaryotic expression vector pcDNA3.1 (-) B/myc-BRMS1 Group receives three monoclonal, Were named as SGC7901-R1, SGC7901-R2, SGC7901-R3; Transfected with empty vector pcDNA3.1/myc-his (-) B group received two monoclonal, Were named as SGC7901-V1, SGC7901-V2; By RT-PCR test after transfection BRMS1 mRNA expression changes; See transfer of eukaryotic expression vector of only three monoclonal SGC7901-R3 has BRMS1 gene expression, Transfer of P group of two monoclonal no BRMS1 gene expressio; Will SGC7901, SGC7901-V1, SGC7901-R3 cells were drawn by MTT method of growth curve, The results show that: BRMS1 gene does not affect gastric cancer cell proliferation SGC7901; Boyden chamber with improved lines of cells in vitro invasion and metastasis experiment, The results showed that: SGC7901-R3 group than SGC7901-V1 group significantly reduced the number of cell invasion, The results are significant differences (P <0.05); SGC7901-V1 group and the group SGC7901, Invasion of no significant reduction in volume, results no significant difference (P> 0.05).Conclusion:BRMS1 gene in vitro SGC7901 gastric cancer cells can inhibit the invasion, but it does not affect their proliferation.