Effects of eukaryotic expression vector pcDNA3.1-Annexin A1 on migration and invasion of colon cancer cells

Abstract

Objective To evaluate the effects of Annexin A1 on migratory and invasive ability of colon cancer cells.Methods Eukaryotic expression vector pcDNA3.1-Annexin A1 was constructed and analyzed by restriction analysis and sequencing.The recombinant plasmid pcDNA3.1-Annexin A1 was transfected into SW480 cells by liposome-mediated gene transferred method.The stable transfectants were obtained after screening with G418.Real-time PCR and Western blotting analysis were used to examine the expression of Annexin A1 mRNA and protein in SW480 cells before and after transfection.Wound-healing experiment and Transwell invasion assays were used to study the effects of Annexin A1 on the migratory and invasive ability of SW480 cells.Results The recombinant plasmid pcDNA3.1-Annexin A1 was successfully constructed.The results of real-time PCR and Western blotting analysis showed that the Annexin A1 expression was significantly higher in cells transfected with pcDNA3.1-Annexin A1 than in un-transfected cells or those transfected with empty vectors(P0.01),while there was no significant difference between the last two groups(P0.05).The migratory rate of SW480 cells in pcDNA3.1-Annexin A1 group was significantly higher than those in un-transfected cells or transfected with empty vectors(\[0.415±0.002) vs \[0.267±0.003\] and \[0.271±0.002\],P0.05),and there was no significant difference between the latter two groups.The migrating SW480 cells in pcDNA3.1-Annexin A1 group was significantly more than those in the other two groups(\[221.75±12.07\] vs \[162.80±12.07\] and \[164.25±9.50\],P0.05).Conclusion Overexpression of Annexin A1 can increase the migratory and invasive ability of SW480 cell line.