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Regulation of the p21Sdi1/Cip1/Waf1 DNA Synthesis Inhibitor in Senescent Human Diploid Fibroblasts

Published online by Cambridge University Press:  29 November 2010

Ryan S. Robetorye  and
James R. Smith
Ryan S. Robetorye
Affiliation:
Baylor College of Medicine
James R. Smith
Affiliation:
Baylor College of Medicine
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Abstract

A large body of evidence has demonstrated that normal human fibroblasts have a limited division potential in culture and underwent senescence, a process whereby cells became arrested in the G1 phase of the cell cycle and overexpressed a DNA synthesis inhibitor(s). Cyclin-dependent kinase two (Cdk2) is required for the promotion of the Gi-to-S phase transition in human cells. Senescent fibroblasts contain intact cyclin-Cdk2 complexes but cannot induce Cdk2 protein kinase activity in response to mitogen stimulation. Recently, we cloned p21Sdi1, a potent inhibitor of DNA synthesis and Cdk2 kinase activity, from a senescent cell cDNA library and demonstrated that it was expressed at significantly higher levels in senescent cells than actively proliferating cells. In contrast to actively dividing cells, mitogen-stimulated senescent cells do not down-regulate the expression of p21Sdi1 and do not express late G1 phase gene products that are required for entry into S phase. We suggest that the inability of mitogen-stimulated senescent cells to down-regulate p21Sdi1 levels contributes to the resulting lack of late Gi gene expression and failure to traverse the G1/S phase boundary.

Résumé

Il a été abondamment démontré que les fibroblastes humains normaux possèdent, en culture, un potentiel de division limité et sont soumis à la sénescence, un processus par lequel les cellules sont arrêtées à l'étape G1 du cycle cellulaire et contiennent une surabondance d'inhibiteurs de synthèse d'ADN. Le passage de la cellule humaine de l'étape G1 à l'étape S nécessite une kinase deux qui dépend de la cycline (Cdk2). Les fibroblastes en sénescence contiennent des complexes de Cdk2 mais ne peuvent produire l'activité de la protéine kinase Cdk2 en réaction à une stimulation mitogène. Récemment, à partir d'une banque D'ADNc de cellules sénescentes, nous avons cloné le p21Sdi1, un inhibiteur potentiel de la réplication de l'ADN et de l'activité de la kinase Cdk2, et démontré qu'on pouvait le trouver à des niveaux beaucoup plus élevés dans les cellules sénescentes que dans les cellules proliférantes actives. À l'encontre des cellules actives en division, les cellules sénescentes ne contrôlent pas à la baisse l'expression du p21Sdi1 et ne produisent pas de produits génétiques en phase G1 avancée nécessaires au passage en phase S. Nous croyons que l'incapacité des cellules sénescentes stimulées par mitogène de contrôler à la baisse les niveaux de p21Sdi1 contribue au manque d'expression génétique de la phase G1 et à l'échec du passage de la phase G1 à la phase S.

Keywords

Cell CycleDNA SynthesisCdk2Cdk Inhibitorp21Sdi1.Cycle cellulairesynthèse d'ADNCdk2inhibiteur de Cdk, p21Sdi1.
Type
Articles
Information
Canadian Journal on Aging / La Revue canadienne du vieillissement , Volume 15 , Issue 2 , Été/Summer 1996 , pp. 315 - 329
Copyright
Copyright © Canadian Association on Gerontology 1996

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