Construction and expression of the recombinant plasmid BFP-cyclin D1 and its influence on proliferation and migration in breast cancer cells MCF-7

Objective To construct the eukaryotic expression plasmid BFP-cyclin D1 and investigate the effects of cyclin D1 on the proliferation and migration of MCF-7 cells.Methods The cyclin D1 gene was amplified by RT-PCR from total RNA of MCF-7 cells,digested with EcoR I and Sal I,and inserted into the eukaryotic florescence expression vector pEBFP-N1.The resulting recombinant plasmid BFP-cyclin D1 was transfected into MCF-7 cells.In parallel,one negative control group was transfected with pEBFP-N1 and another with transfection reagent without any DNA.Following transfection,cell proliferation was detected by MTT assay,cell migration by cell scratch assay,and mRNA expression by quantitative real-time PCR.Results The eukaryotic expression plasmid BFP-cyclin D1 was successfully constructed and high expression of cyclin D1 was detected in MCF-7 cells.Cell proliferation and migration were significantly higher in the cells transfected with BFP-cyclin D1 than in the two control groups(P0.01).Conclusions High expression of cyclin D1 can accelerate proliferation and migration of MCF-7 cells,perhaps by shortening the cell cycle and promoting DNA replication.