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.
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More From: Academic Journal of Second Military Medical University
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