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Cell Growth & Differentiation, Vol 9, Issue 1 85-95, Copyright © 1998 by American Association of Cancer Research


ARTICLES

Replicative senescence of normal human oral keratinocytes is associated with the loss of telomerase activity without shortening of telomeres

MK Kang, W Guo and NH Park
Dental Research Institute, School of Dentistry, and Jonsson Comprehensive Cancer Center, University of California, Los Angeles 90095-1668, USA.

Telomerase activity was analyzed in 7 different cultures of secondary normal human oral keratinocytes (NHOKs), 1 normal human oral epithelial tissue specimen, 1 immortalized human oral keratinocyte (HOK) cell line, and 10 human oral cancer cell lines using the PCR-based telomeric repeat amplification protocol assay. Telomerase activity was found in all tested cells and tissue, but the activity in NHOKs and epithelial tissue was lower than that in other tested cell lines. Inasmuch as continued subculture of NHOKs results in replicative senescence, we investigated the association between telomerase activity and replicative senescence by evaluating the enzyme activity in NHOK cultures with different population doubling levels. Three different NHOK cultures were independently subcultured until these cells reached the postmitotic stage. Unlike in fibroblasts derived from the human oral cavity, significant telomerase activity was detected in rapidly proliferating NHOKs, and telomerase activity was barely detectable in the keratinocytes near and at senescence. However, the terminal restriction fragment consisting of telomeric DNA was found to be constantly maintained at approximately 6.0 kilobases in NHOKs without any detectable shortening of telomeres by subcultures. Intracellular p53 and p21WAF1/CIP1 protein levels in NHOKs were gradually and significantly diminished by the passage of cells. These data indicate that actively proliferating NHOKs contain telomerase activity and that replicative senescence of NHOKs is associated with the loss of telomerase activity without shortening of telomeres. However, replicative senescence of NHOKs is apparently not linked to an accumulation of wild-type p53 and/or p21WAF1/CIP1 proteins in these cells.


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Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cell Growth & Differentiation
Copyright © 1998 by the American Association of Cancer Research.