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Concurrence of replicative senescence and elevated expression of p16INK4A with subculture-induced but not calcium-induced differentiation in normal human oral keratinocytes

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Authors
Lee, Gene; Park, Beom Seok; Han, Song Ee; Oh, Ju-Eun; You, Yong-Ouk; Baek, Jeong-Hwa; Kim, Gwan-Shik; Min, Byung-Moo
Issue Date
2000-10
Publisher
Elsevier
Citation
Archives of Oral Biology 45, (2000), 809-818
Keywords
Calcium-induced differentiationSubculture-induced differentiationNormal human oral keratinocytesReplicative senesencep16INK4A
Abstract
Primary normal human oral keratinocytes (NHOKs) undergo differentiation in the presence of calcium concentrations higher than 0.15 mM in vitro, which is useful in investigating the mechanisms involved in the differentiation of epithelial cells. Serial subculture of NHOKs to the postmitotic stage also induces terminal differentiation. However, the detailed mechanisms of both differentiation processes remain substantially unknown. To investigate the molecular differences in these processes, NHOKs were induced to differentiate by exposure to 1.2 mM of calcium and by serial subculture to the postmitotic stage. To study whether the cells were induced to differentiate and to undergo replicative senescence, the amount of cellular involucrin and the expression of senescence-associated β-galactosidase (SA-β-gal) were measured respectively. The expression of replicative senescence-associated genes and the activity of telomerase from the differentiated cells were also determined. Both calcium treatment and serial subculture to the postmitotic stage notably elevated the cellular involucrin. The percentage of SA-β-gal-positive cells was significantly elevated by the continued subculture, but such changes were not observed in keratinocytes exposed to calcium. The concentration of cellular p16INK4A protein was progressively increased by the continued subculture but was not changed by calcium treatment. On the other hand, the concentrations of cellular p53 were similar in both differentiation processes. However, telomerase activity was lost in NHOKs that had undergone differentiation by both calcium treatment and serial subculture. The results indicate that calcium-induced differentiation of NHOKs has similar characteristics to their serial subculture-induced differentiation, but that the differentiation processes are not identical, because calcium-induced differentiation does not concur with either replicative senescence or the gradually increased concentration of p16INK4A.
ISSN
0003-9969
Language
English
URI
http://hdl.handle.net/10371/63035
DOI
https://doi.org/10.1016/S0003-9969(00)00061-3
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College of Dentistry/School of Dentistry (치과대학/치의학대학원)Dept. of Dentistry (치의학과)Journal Papers (저널논문_치의학과)
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