Senescence delay of human diploid fibroblast induced by anti-sense p16INK4a expression.
p16(INK4a), a tumor suppressor gene that inhibits cyclin-dependent kinase 4 and cyclin-dependent kinase 6, is also implicated in the mechanisms underlying replicative senescence, because its RNA and protein accumulate as cells approach their finite number of population doublings in tissue culture. To further explore the involvement of p16(INK4a) in replicative senescence, we constructed a retroviral vector containing antisense p16(INK4a), pDOR-ASp16, and introduced it into early passages of human diploid fibroblasts. The introduction of this construct significantly suppressed the expression of wild-type p16(INK4a). It also imposed a finite increase in proliferative life span and significant delay of several other cell senescent features, such as cell flattening, cell cycle arrest, and senescence-associated beta-galactosidase positivity. Moreover, telomere shortening and decline in DNA repair capacity, which normally accompany cell senescence, are also postponed by the ASp16 transfection. The life span of fibroblasts was significantly extended, but the onset of replicative senescence could not be totally prevented. Telomerase could not be activated even though telomere shortening was slowed. These observations suggest that the telomere pathway of senescence cannot be bypassed by ASp16 expression. These data not only strongly support a role for p16(INK4a) in replicative senescence but also raise the possibility of using the antisense p16(INK4a) therapeutically.
Duan
J
J
Department of Biochemistry and Molecular Biology, Health Science Center, Peking University, Beijing 100083, People's Republic of China.
Zhang
Z
Z
Tong
T
T
D001483:Base Sequence; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000235:genetics*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D017931:DNA Primers; D004260:DNA Repair; D004171:Diploidy; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D019941:Cyclin-Dependent Kinase Inhibitor p16; D017931:DNA Primers; D016376:Oligonucleotides, Antisense; D001616:beta-Galactosidase