Construction Of P27 Kip1 Eukaryotic Expression Plasmid To Investigate Its Influence On Mononuclear Cell Cycle

Aim Constructing p27kip1 eukaryotic expression plasmid to investigate its influence on mononuclear cell proliferation and cell cycle for further researching the role of p27kip1 in disease prevention and treatment. Methods p27kip1 gene was cloned into pcDNA3.0 eukaryotic expression, then pcDNA3.0/ p27kip1 mediated by liposome was transfected into mononuclear cell. MTT assay and flow cytometry were used to analyze cell proliferation and cell cycle. Result We successfully constructed the mouse pcDNA3.0/p27kip1 eukaryotic expression plasmid. The cells transfected with pcDNA3.0/ p27kip1 expressed more p27 protein, and their proliferation was inhibited obviously. The percentage of the cells in G0/G1 phase increased, and that in S phase decreased significantly. Conclusion Mouse pcDNA3.0/ p27kip1 eukaryotic expression plasmid can be expressed in mononuclear cells, and overexpression of p27 protein can inhibit mononuclear cell proliferation by restricting the cell cycle.