Construction of p27~(kip1) eukaryotic expression plasmid to investigate its influence on mononuclear cell cycle
- VernacularTitle:小鼠p27~(kip1)真核表达载体的构建及其对单个核细胞周期的影响
- Author:
Wei ZHU
;
Ping CHEN
;
Haitao ZHAO
;
Li SUN
- Publication Type:Journal Article
- Keywords:
p27~(kip1);
mononuclear cell;
cell cycle
- From:
Chinese Pharmacological Bulletin
2003;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Aim To construct 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.Results The mouse pcDNA3.0/p27kip1 eukaryotic expression plasmid were successfully constructed.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.Conclusions Mouse pcDNA3.0/p27kip1 eukaryotic expression plasmid could be expressed in mononuclear cells,and overexpression of p27 protein could inhibit mononuclear cell proliferation by restricting the cell cycle,making cell cycle arrest in G0/G1 phase.
