title: Effect of subcellular localization of P21 on proliferation and apoptosis of HepG2 cells.
abstract: This study examined the effect of subcellular localization of P21 on the proliferation and apoptosis of HepG2 cells. The coding genes of the wild and the mutant P21 were amplified by mega primer PCR from the plasmid pCEP-WAF1 which contains human P21 cDNA in the nuclear localizational signal (NLS) sequence, and then inserted into the eukaryotic expression vector pDsRed1-C1. The recombinants were transfected into HepG2 cells. The transcription and expression of P21 were determined by RT-PCR and fluorescence microscopy. The cell proliferation was measured by MTT, and the cell cycle and apoptosis of HepG2 cells by flow cytometry. The results of restriction analysis, DNA sequencing and fluorescence microscopy confirmed the construction of the wild and the mutant P21 in the eukaryotic expression plasmid. The plasmid containing the mutant P21 was found to accelerate cell proliferation and the wild P21 plasmid to inhibit cell proliferation. Cell cycle analysis showed that the cell ratio of G(0)/G(1) in the wild type group was significantly increased as compared with that in the mutant type group, and cell apoptosis analysis revealed that the apoptosis rate in the wild type group was much higher than that in the mutant type group. It was concluded that the subcellular localization of P21 may contribute to the development of hepatic cancer.
authors.affiliation: Department of Gastroenterology, Yueyang People's Hospital, Hunan Normal University, Yueyang, 414000, China.|Department of Hepatic Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
authors.forename: Rongyuan
authors.initials: R
authors.lastname: Qiu
authors.affiliation: Department of Gastroenterology, Yueyang People's Hospital, Hunan Normal University, Yueyang, 414000, China.
authors.forename: Songbai
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology, Yueyang People's Hospital, Hunan Normal University, Yueyang, 414000, China.
authors.forename: Xihua
authors.initials: X
authors.lastname: Feng
authors.affiliation: Department of Hepatic Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Chen
authors.affiliation: Department of Hepatic Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
authors.forename: Kaikai
authors.initials: K
authors.lastname: Yang
authors.affiliation: Department of Hepatic Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. shengshe168@yahoo.com.cn.
authors.forename: Shengsong
authors.initials: S
authors.lastname: He
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism* / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism*; D056945:Hep G2 Cells; D006801:Humans; D008969:Molecular Sequence Data; D014018:Tissue Distribution
chemical_list: C493956:CDKN1A protein, human; D050759:Cyclin-Dependent Kinase Inhibitor p21