Effects of eukaryotic expression vector pcDNA3.1-Annexin A1 on migration and invasion of colon cancer cells
10.3724/SP.J.1008.2010.00264
- Author:
Xiao-Qun YANG
1
Author Information
1. Department of Pathology
- Publication Type:Journal Article
- Keywords:
Annexin A1 gene;
Colorectal neoplasms;
Eukaryotic expression;
Transwell invasion assays;
Wound-healing experiment
- From:
Academic Journal of Second Military Medical University
2010;31(3):264-268
- CountryChina
- Language:Chinese
-
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 (P<0. 01), while there was no significant difference between the last two groups (P>0.05). 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], P<0.05), 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], P<0. 05). Conclusion Overexpression of Annexin A1 can increase the migratory and invasive ability of SW480 cell line.
