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Features of replicative senescence induced by direct addition of antennapedia‐p16 INK4A fusion protein to human diploid fibroblasts
Author(s) -
Kato Daishiro,
Miyazawa Kazuhiro,
Ruas Marugarida,
Starborg Maria,
Wada Ikuo,
Oka Takahiro,
Sakai Toshiyuki,
Peters Gordon,
Hara Eiji
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00426-8
Subject(s) - antennapedia , biology , senescence , fusion protein , cyclin dependent kinase , retinoblastoma protein , population , transfection , microbiology and biotechnology , phenotype , protein kinase a , kinase , cell culture , cell cycle , genetics , cell , gene , recombinant dna , homeotic gene , demography , sociology
The p16 INK4A cyclin‐dependent kinase (Cdk) inhibitor is now recognized as a major tumor suppressor that is inactivated by a variety of mechanisms in a wide range of human cancers. It is also implicated in the mechanisms underlying replicative senescence since p16 INK4A RNA and protein accumulate as cells approach their proscribed limit of population doublings in tissue culture. To obtain further evidence of its role in senescence, we have sought ways of overexpressing p16 INK4A in primary human diploid fibroblasts (HDF). To circumvent the low transfection efficiency of primary cells we have exploited a recombinant form of the full‐length p16 INK4A protein fused to a 16 amino acid peptide from the Drosophila antennapedia protein. This peptide has the capacity to cross both cytoplasmic and nuclear membranes allowing the direct introduction of the active protein to primary cells. Here, we show that antennapedia‐tagged wild‐type p16 INK4A protein, but not a functionally compromised tumor‐specific variant, causes G1 arrest in early passage HDFs by inhibiting the phosphorylation of the retinoblastoma protein. Significantly, the arrested cells display several phenotypic features that are considered characteristic of senescent cells. These data support a role for p16 INK4A in replicative senescence and raise the possibility of using the antennapedia‐tagged protein therapeutically.