Effect of cell cycle inhibitor p19ARF on senescence of human diploid cell.
To investigate the effect of cell cycle inhibitor p14ARF on replicative senescence of human diploid cell, recombinant p19ARF eukaryotic expression vector was constructed and p19ARF gene was transfected into human diploid fibroblasts (WI-38 cells) by liposome-mediated transfection for overexpression. Then, the effects of p19ARF on replicative senescence of WI-38 cells were observed. The results revealed that, compared with control cells, the WI-38 cells in which p19ARF gene was introduced showed significant up-regulation of p53 and p21 expression level, decrease of cell generation by 10-12 generations, decline of cell growth rate with cell cycle being arrested at G1 phase, increase of positive rate of senescent marker SA-beta-gal staining, and decrease of mitochondrial membrane potential. The morphology of the transfected fibroblasts presented the characteristics changes similar to senescent cells. These results indicated that high expression of p19ARF may promote the senescent process of human diploid cells.
Bai
XueYuan
X
Chinese PLA Institute of Nephrology, Chinese PLA General Hospital & Military Medical Postgraduate College, Beijing 100853, China. baixy@301hospital.com.cn
Chen
XiangMei
X
Hou
Kai
K
Zhang
Ping
P
Feng
Zhe
Z
Fu
Bo
B
D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D016922:Cellular Senescence / Q000187:drug effects*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000494:pharmacology*; D004171:Diploidy; D005434:Flow Cytometry; D016158:Genes, p53; D005822:Genetic Vectors; D006801:Humans; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D014162:Transfection; D054462:p21-Activated Kinases
D019941:Cyclin-Dependent Kinase Inhibitor p16; C515903:PAK1 protein, human; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases