| Objective:Construction a BBS1 eukaryotic expression vector to explore whether BBS1 gene effects on cell proliferation and migration;To study the correlation among BBS1 and Annexin A1,kif2a,KIF3A,β-Tublin,β-catenin.Method:A commercial BBS1 eukaryotic expression plasmid was used in this study.BBS1 high-level and low-level expression cell models were constructed by transient transfection/cotransfection the BBS1 eukaryotic expression vector and it’s siRNA vectors,respectivel.MTS experimental and cell scratch experimental were used to test whether cell proliferation and migration were effected by BBS1 gene,respectively.Total RNA and proteins of transfection cells were extracted,BBSI level was detedcted by using qPCR and western blot.Relationship among BBS 1 and Annexin A1,kif2a,kif3a,β-Tublin,β-catenin were analaysed by using SPSS software.Result:BBS1 and it’s siRNAs were successfully transfected into the 298T cell and were positive in PCR and western blotting;High-level of BBS1 gene might inhibit the proliferation and migration of 293T cell;Annexin A1,kif2a,KIF3A,β-Tublin,β-catenin were positively correlation with BBS1,such as,in the siRNA 1 interference group,KIF3A gene expression was increasing with the high level of BBS1,and the expression of Kif2a and ANXA1 were consistent with the BBSI level.Conclusion:The relationship among BBS1 and Annexin Al,kif2a,KIF3A might give new clues to explore the mechanism of Bardet Biedl Syndrome.The effect of high-level BBS1 on cell proliferation might give positive contribution to cancer therapy. |
